Phosphate compounds, a process for their preparation and pharmaceutical compositions containing them

ABSTRACT

Compounds of formula (I): 
     
       
         
         
             
             
         
       
     
     wherein X, Y, A 1 , A 2 , R a , R b , R c , R d , R 3 , R 4 , T and R 5  are as defined in the description. Medicinal products containing the same which are useful in treating pathologies involving a deficit in apoptosis, such as cancer, auto-immune diseases, and diseases of the immune system.

The present invention relates to new phosphate compounds, to a processfor their preparation and to pharmaceutical compositions containingthem.

The compounds of the present invention are new and have very valuablepharmacological and pharmacokinetic characteristics for use in the fieldof apoptosis and cancerology.

Apoptosis, or programmed cell death, is a physiological process that iscrucial for embryonic development and maintenance of tissue homeostasis.

Apoptotic-type cell death involves morphological changes such ascondensation of the nucleus, DNA fragmentation and also biochemicalphenomena such as the activation of caspases which cause damage to keystructural components of the cell, so inducing its disassembly anddeath. Regulation of the process of apoptosis is complex and involvesthe activation or repression of several intracellular signaling pathways(Cory S. et al., Nature Review Cancer, 2002, 2, 647-656).

Deregulation of apoptosis is involved in certain pathologies. Increasedapoptosis is associated with neurodegenerative diseases such asParkinson's disease, Alzheimer's disease and ischaemia. Conversely,deficits in the implementation of apoptosis play a significant role inthe development of cancers and their chemoresistance, in auto-immunediseases, inflammatory diseases and viral infections. Accordingly,absence of apoptosis is one of the phenotypic signatures of cancer(Hanahan D. et al., Cell 2000, 100, 57-70).

The anti-apoptotic proteins of the Bcl-2 family are associated withnumerous pathologies. The involvement of proteins of the Bcl-2 family isdescribed in numerous types of cancer, such as colorectal cancer, breastcancer, small-cell lung cancer, non-small-cell lung cancer, bladdercancer, ovarian cancer, prostate cancer, chronic lymphoid leukaemia,follicular lymphoma, myeloma, etc. Overexpression of the anti-apoptoticproteins of the Bcl-2 family is involved in tumorigenesis, in resistanceto chemotherapy and in the clinical prognosis of patients affected bycancer. There is, therefore, a therapeutic need for compounds thatinhibit the anti-apoptotic activity of the proteins of the Bcl-2 family.

In addition to being new, the compounds of the present invention havepharmacological and pharmacokinetic properties making it possible to usethem in pathologies involving a defect in apoptosis, such as, forexample, in the treatment of cancer, auto-immune diseases and diseasesof the immune system.

The present invention relates more especially to a phosphate compound offormula (I):

wherein:

-   -   X and Y represent a carbon atom or a nitrogen atom, it being        understood that they may not simultaneously represent two        carbons atoms or two nitrogen atoms,    -   A₁ and A₂, together with the atoms carrying them, form an        optionally substituted, aromatic or non-aromatic heterocycle Het        composed of 5, 6 or 7 ring members which may contain, in        addition to the nitrogen represented by X or by Y, from one to 3        hetero atoms selected independently from oxygen, sulphur and        nitrogen, it being understood that the nitrogen in question may        be substituted by a group representing a hydrogen atom, a linear        or branched (C₁-C₆)alkyl group or a group —C(O)—O-Alk wherein        Alk is a linear or branched (C₁-C₆)alkyl group,    -   or A₁ and A₂ independently of one another represent a hydrogen        atom, a linear or branched (C₁-C₆)polyhaloalkyl, a linear or        branched (C₁-C₆)alkyl group or a cycloalkyl,    -   T represents a hydrogen atom, a linear or branched (C₁-C₆)alkyl        group optionally substituted by from one to three halogen atoms,        a group (C₁-C₄)alkyl-NR₁R₂, or a group (C₁-C₄)alkyl-OR₆,    -   R₁ and R₂ independently of one another represent a hydrogen atom        or a linear or branched (C₁-C₆)alkyl group,    -   or R₁ and R₂ form with the nitrogen atom carrying them a        heterocycloalkyl,    -   R₃ represents a linear or branched (C₁-C₆)alkyl group, a linear        or branched (C₂-C₆)alkenyl group, a linear or branched        (C₂-C₆)alkynyl group, a cycloalkyl group, a        (C₃-C₁₀)cycloalkyl-(C₁-C₆)alkyl group wherein the alkyl moiety        is linear or branched, a heterocycloalkyl group, an aryl group        or a heteroaryl group, it being understood that one or more of        the carbon atoms of the preceding groups, or of their possible        substituents, may be deuterated,    -   R₄ represents an aryl group, a heteroaryl group, a cycloalkyl        group or a linear or branched (C₁-C₆)alkyl group, it being        understood that one or more of the carbon atoms of the preceding        groups, or of their possible substituents, may be deuterated,    -   R₅ represents a hydrogen or halogen atom, a linear or branched        (C₁-C₆)alkyl group, or a linear or branched (C₁-C₆)alkoxy group,    -   R₆ represents a hydrogen atom or a linear or branched        (C₁-C₆)alkyl group,    -   R_(a), R_(b), R_(c) and R_(d), each independently of the others,        represent R₇, a halogen atom, a linear or branched (C₁-C₆)alkoxy        group, a hydroxy group, a linear or branched        (C₁-C₆)polyhaloalkyl group, a trifluoromethoxy group, —NR₇R₇′,        nitro, R₇—CO—(C₀-C₆)alkyl-, R₇—CO—NH—(C₀-C₆)alkyl-,        NR₇R₇′—CO—(C₀-C₆)alkyl-, NR₇R₇′—CO—(C₀-C₆)alkyl-O—,        R₇—SO₂—NH-—(C₀-C₆)alkyl-, R₇—NH—CO—NH—(C₀-C₆)alkyl-,        R₇—O—CO—NH—(C₀-C₆)alkyl-, a heterocycloalkyl group, or the        substituents of one of the pairs (R_(a),R_(b)), (R_(b),R_(c)) or        (R_(c),R_(d)) form together with the carbon atoms carrying them        a ring composed of from 5 to 7 ring members, which may contain        from one to 2 hetero atoms selected from oxygen and sulphur, it        also being understood that one or more carbon atoms of the ring        defined hereinbefore may be deuterated or substituted by from        one to 3 groups selected from halogen and linear or branched        (C₁-C₆)alkyl,    -   R₇ and R₇′ independently of one another represent a hydrogen, a        linear or branched (C₁-C₆)alkyl, a linear or branched        (C₂-C₆)alkenyl, a linear or branched (C₂-C₆)alkynyl, an aryl or        a heteroaryl, or R₇ and R₇′ together with nitrogen atom carrying        them form a heterocycle composed of from 5 to 7 ring members,

the compound of formula (I) being such that at least one of the carbonatoms contained in it is substituted by one of the following phosphategroups: —OPO(OM)(OM′), —OPO(OM)(O⁻M₁ ⁺), —OPO(O⁻M₁ ⁺)(O⁻M₂ ⁺),—OPO(O⁻)(O⁻)M₃ ²⁺, —OPO(OM)(O[CH₂CH₂O]_(n)CH₃), or —OPO(O⁻M₁⁺)(O[CH₂CH₂O]_(n)CH₃), wherein M and M′ independently of one anotherrepresent a hydrogen atom, a linear or branched (C₁-C₆)alkyl group, alinear or branched (C₂-C₆)alkenyl group, a linear or branched(C₂-C₆)alkynyl group, a cycloalkyl or a heterocycloalkyl both composedof 5 or 6 ring members, while M₁ ⁺ and M₂ ⁺ independently of one anotherrepresent a pharmaceutically acceptable monovalent cation, and M₃ ²⁺represents a pharmaceutically acceptable divalent cation and n is aninteger from 1 to 5,

it being understood that:

-   -   “aryl” means a phenyl, naphthyl, biphenyl or indenyl group,    -   “heteroaryl” means any mono- or bi-cyclic group composed of from        5 to 10 ring members, having at least one aromatic moiety and        containing from 1 to 4 hetero atoms selected from oxygen,        sulphur and nitrogen (including quaternary nitrogens),    -   “cycloalkyl” means any mono- or bi-cyclic, non-aromatic,        carbocyclic group containing from 3 to 10 ring members,    -   “heterocycloalkyl” means any mono- or bi-cyclic, non-aromatic,        condensed or spiro group composed of 3 to 10 ring members and        containing from 1 to 3 hetero atoms selected from oxygen,        sulphur, SO, SO₂ and nitrogen,

it being possible for the aryl, heteroaryl, cycloalkyl andheterocycloalkyl groups so defined and the groups alkyl, alkenyl,alkynyl and alkoxy to be substituted by from 1 to 3 groups selected fromoptionally substituted, linear or branched (C₁-C₆)alkyl, (C₃-C₆)spiro,linear or branched, optionally substituted (C₁-C₆)alkoxy,(C₁-C₆)alkyl-S—, hydroxy, oxo (or N-oxide where appropriate), nitro,cyano, —COOR′, —OCOR′, NR′R″, linear or branched (C₁-C₆)polyhalo alkyl,trifluoromethoxy, (C₁-C₆)alkylsulphonyl, halogen, optionally substitutedaryl, heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyloptionally substituted by one or more halogen atoms or alkyl groups, itbeing understood that R′ and R″ independently of one another represent ahydrogen atom or an optionally substituted, linear or branched(C₁-C₆)alkyl group,

it being possible for the Het group defined in formula (I) to besubstituted by from one to three groups selected from linear or branched(C₁-C₆)alkyl, hydroxy, linear or branched (C₁-C₆)alkoxy, NR₁′R₁″ andhalogen, it being understood that R₁′ and R₁″ are as defined for thegroups R′ and R″ mentioned hereinbefore,

to its enantiomers and diastereoisomers, and to addition salts thereofwith a pharmaceutically acceptable acid or base.

Among the pharmaceutically acceptable acids there may be mentioned,without implying any limitation, hydrochloric acid, hydrobromic acid,sulphuric acid, phosphonic acid, acetic acid, trifluoroacetic acid,lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid,fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid,oxalic acid, methanesulphonic acid, camphoric acid etc.

Among the pharmaceutically acceptable bases there may be mentioned,without implying any limitation, sodium hydroxide, potassium hydroxide,triethylamine, tert-butylamine etc.

Preferred compounds of the invention include compounds of formula (I)wherein R₄ represents phenyl substituted in the para position by a groupof formula —OPO(OM)(OM′), —OPO(OM)(O⁻M₁ ⁺), —OPO(O⁻M₁ ⁺)(O⁻M₂ ⁺),—OPO(O⁻)(O⁻)M₃ ²⁺, —OPO(OM)(O[CH₂CH₂O]_(n)CH₃), or —OPO(O⁻M₁⁺)(O[CH₂CH₂O]_(n)CH₃), wherein M and M′ independently of one anotherrepresent a hydrogen atom, a linear or branched (C₁-C₆)alkyl group, alinear or branched (C₂-C₆)alkenyl group, a linear or branched(C₂-C₆)alkynyl group, a cycloalkyl or a heterocycloalkyl both composedof 5 or 6 ring members, while M₁ ⁺ and M₂ ⁺ independently of one anotherrepresent a pharmaceutically acceptable monovalent cation, and M₃ ²⁺represents a pharmaceutically acceptable divalent cation and n is aninteger from 1 to 5, it being understood that the phenyl group mayoptionally be substituted by one or more halogen atoms.

Preference is given to compounds of formula (I) wherein R₄ represents aphenyl or a pyrimidin-5-yl group, both substituted in the para positionby a group of formula —OPO(O⁻M₁ ⁺)(O⁻M₂ ⁺), and even more especially bya group of formula —OPO(O⁻Na⁺)(O⁻Na⁺).

Advantageously, X represents a carbon atom and Y represents a nitrogenatom. Even more advantageously, the group:

represents a 5,6,7,8-tetrahydroindolizine, an indolizine or adimethylated pyrrole.

T preferably represents a methyl, (morpholin-4-yl)methyl or3-(morpholin-4-yl)propyl group.

In preferred compounds of the invention, R_(a) and R_(d) each representa hydrogen atom and (R_(b),R_(c)), together with the carbon atomscarrying them, form a 1,3-dioxolane group or a 1,4-dioxane group; orR_(a), R_(c) and R_(d) each represent a hydrogen atom and R_(b)represents a hydrogen or a halogen.

In another embodiment of the invention, R_(a) and R_(d) each represent ahydrogen atom, R_(b) represents a halogen atom and R_(c) a methoxygroup.

Alternatively, R_(a), R_(b) and R_(d) each advantageously represent ahydrogen atom and R_(c) represents a group NR₇R₇′—CO—(C₀-C₆)alkyl-O—,and even more preferably R_(c) represents a2-oxo-2-(piperidin-1-yl)ethoxy group.

Furthermore, R₃ advantageously represents a group selected from phenyl,1H-indole, 1H-pyrrolo[2,3-b]pyridine, pyridine, 1H-pyrazole, 1H-pyrroleand 2,3-dihydro-1H-pyrrolo[2,3-b]pyridine, those groups optionallyhaving one or more substituents selected from linear or branched(C₁-C₆)alkyl (more preferably methyl), cyano and trideuteriomethyl.

Among the preferred compounds of the invention there may be mentioned:

-   -   4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyl        disodium phosphate,    -   4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyl        disodium phosphate,    -   4-({[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideuteriomethyl)-1H-pyrazol-4-yl]amino)phenyl        disodium phosphate,    -   4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)amino]phenyl        disodium phosphate,    -   4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-methyl-1H-pyrrol-3-yl)amino]phenyl        disodium phosphate,    -   4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyl        disodium phosphate,    -   4-[(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}amino]phenyl        disodium phosphate,    -   4-[{[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyl        disodium phosphate,

their enantiomers and diastereoisomers, and addition salts thereof witha pharmaceutically acceptable acid or base.

Pharmacokinetic study of the phosphate compounds of formula (I) showedthat they were converted in vivo into compounds of formula (I′)characterised in that the phosphate function was metabolised into ahydroxy function. The compounds of formula (I) accordingly behave asprodrugs of compounds of formula (I′) having the following formula:

wherein:

-   -   X and Y represent a carbon atom or a nitrogen atom, it being        understood that they may not simultaneously represent two        carbons atoms or two nitrogen atoms,    -   A₁ and A₂, together with the atoms carrying them, form an        optionally substituted, aromatic or non-aromatic heterocycle Het        composed of 5, 6 or 7 ring members which may contain, in        addition to the nitrogen represented by X or by Y, from one to 3        hetero atoms selected independently from oxygen, sulphur and        nitrogen, it being understood that the nitrogen in question may        be substituted by a group representing a hydrogen atom, a linear        or branched (C₁-C₆)alkyl group or a group —C(O)—O-Alk wherein        Alk is a linear or branched (C₁-C₆)alkyl group,    -   or A₁ and A₂ independently of one another represent a hydrogen        atom, a linear or branched (C₁-C₆)polyhaloalkyl, a linear or        branched (C₁-C₆)alkyl group or a cycloalkyl,    -   T represents a hydrogen atom, a linear or branched (C₁-C₆)alkyl        group optionally substituted by from one to three halogen atoms,        a group (C₁-C₄)alkyl-NR₁R₂, or a group (C₁-C₄)alkyl-OR₆,    -   R₁ and R₂ independently of one another represent a hydrogen atom        or a linear or branched (C₁-C₆)alkyl group,    -   or R₁ and R₂ form with the nitrogen atom carrying them a        heterocycloalkyl,    -   R₃ represents a linear or branched (C₁-C₆)alkyl group, a linear        or branched (C₂-C₆)alkenyl group, a linear or branched        (C₂-C₆)alkynyl group, a cycloalkyl group, a        (C₃-C₁₀)cycloalkyl-(C₁-C₆)alkyl group wherein the alkyl moiety        is linear or branched, a heterocycloalkyl group, an aryl group        or a heteroaryl group, it being understood that one or more of        the carbon atoms of the preceding groups, or of their possible        substituents, may be deuterated,    -   R₄ represents an aryl group, a heteroaryl group, a cycloalkyl        group or a linear or branched (C₁-C₆)alkyl group, it being        understood that one or more of the carbon atoms of the preceding        groups, or of their possible substituents, may be deuterated,    -   R₅ represents a hydrogen or halogen atom, a linear or branched        (C₁-C₆)alkyl group, or a linear or branched (C₁-C₆)alkoxy group,    -   R₆ represents a hydrogen atom or a linear or branched        (C₁-C₆)alkyl group,    -   R_(a), R_(b), R_(c) and R_(d), each independently of the others,        represent R₇, a halogen atom, a linear or branched (C₁-C₆)alkoxy        group, a hydroxy group, a linear or branched        (C₁-C₆)polyhaloalkyl group, a trifluoromethoxy group, —NR₇R₇′,        nitro, R₇—CO—(C₀-C₆)alkyl-, R₇—CO—NH—(C₀-C₆)alkyl-,        NR₇R₇′—CO—(C₀-C₆)alkyl-, NR₇R₇′—CO—(C₀-C₆)alkyl-O—,        R₇—SO₂—NH—(C₀-C₆)alkyl-, R₇—NH—CO—NH—(C₀-C₆)alkyl-,        R₇—O—CO—NH—(C₀-C₆)alkyl-, a heterocycloalkyl group, or the        substituents of one of the pairs (R_(a),R_(b)), (R_(b),R_(c)) or        (R_(c),R_(d)) form together with the carbon atoms carrying them        a ring composed of from 5 to 7 ring members, which may contain        from one to 2 hetero atoms selected from oxygen and sulphur, it        also being understood that one or more carbon atoms of the ring        defined hereinbefore may be deuterated or substituted by from        one to 3 groups selected from halogen and linear or branched        (C₁-C₆)alkyl,    -   R₇ and R₇′ independently of one another represent a hydrogen, a        linear or branched (C₁-C₆)alkyl, a linear or branched        (C₂-C₆)alkenyl, a linear or branched (C₂-C₆)alkynyl, an aryl or        a heteroaryl, or R₇ and R₇′ together with nitrogen atom carrying        them form a heterocycle composed of from 5 to 7 ring members,

it being understood that:

-   -   “aryl” means a phenyl, naphthyl, biphenyl or indenyl group,    -   “heteroaryl” means any mono- or bi-cyclic group composed of from        5 to 10 ring members, having at least one aromatic moiety and        containing from 1 to 4 hetero atoms selected from oxygen,        sulphur and nitrogen (including quaternary nitrogens),    -   “cycloalkyl” means any mono- or bi-cyclic, non-aromatic,        carbocyclic group containing from 3 to 10 ring members,    -   “heterocycloalkyl” means any mono- or bi-cyclic, non-aromatic,        condensed or spiro group composed of 3 to 10 ring members and        containing from 1 to 3 hetero atoms selected from oxygen,        sulphur, SO, SO₂ and nitrogen,

it being possible for the aryl, heteroaryl, cycloalkyl andheterocycloalkyl groups so defined and the groups alkyl, alkenyl,alkynyl and alkoxy to be substituted by from 1 to 3 groups selected fromoptionally substituted, linear or branched (C₁-C₆)alkyl, (C₃-C₆)spiro,linear or branched, optionally substituted (C₁-C₆)alkoxy,(C₁-C₆)alkyl-S—, hydroxy, oxo (or N-oxide where appropriate), nitro,cyano, —COOR′, —OCOR′, NR′R″, linear or branched (C₁-C₆)polyhalo alkyl,trifluoromethoxy, (C₁-C₆)alkylsulphonyl, halogen, optionally substitutedaryl, heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyloptionally substituted by one or more halogen atoms or alkyl groups, itbeing understood that R′ and R″ independently of one another represent ahydrogen atom or an optionally substituted, linear or branched(C₁-C₆)alkyl group,

it being possible for the Het group defined in formula (I′) to besubstituted by from one to three groups selected from linear or branched(C₁-C₆)alkyl, hydroxy, linear or branched (C₁-C₆)alkoxy, NR₁′R₁″ andhalogen, it being understood that R₁′ and R₁″ are as defined for thegroups R′ and R″ mentioned hereinbefore,

their enantiomers and diastereoisomers, and addition salts thereof witha pharmaceutically acceptable acid or base.

The compounds of formula (I′) have pro-apoptotic properties and as aresult are of major therapeutic value in the treatment of cancers,auto-immune diseases and diseases of the immune system. In the presentinvention it has been shown that, by administering the phosphatecompounds of formula (I), the in vivo exposure to the compounds offormula (I′) was optimised. The solubility of the compounds of formula(I) is in fact much greater than that of the compounds of formula (I′).Consequently, using the compounds of formula (I) in the manufacture ofpharmaceutical compositions is especially advantageous from the galenicpoint of view.

The invention relates also to a process for the preparation of compoundsof formula (I), which process is characterised in that there is used asstarting material the compound of formula (II):

wherein R_(a), R_(b), R_(c) and R_(d) are as defined for formula (I′),

which compound of formula (II) is subjected to a Heck reaction, in anaqueous or organic medium, in the presence of a palladium catalyst, of abase, of a phosphine and of the compound of formula (III):

wherein the groups A₁, A₂, X and Y are as defined for formula (I′) andAlk represents a linear or branched (C₁-C₆)alkyl,

to obtain the compound of formula (IV):

wherein A₁, A₂, X, Y, R_(a), R_(b), R_(c) and R_(d) are as defined forformula (I′) and Alk is as defined hereinbefore,

the aldehyde function of which compound of formula (IV) is oxidised to acarboxylic acid to form the compound of formula (V):

wherein A₁, A₂, X, Y, R_(a), R_(b), R_(c) and R_(d) are as defined forformula (I′) and Alk is as defined hereinbefore,

which compound of formula (V) is then subjected to peptide coupling witha compound of formula (VI):

wherein T and R₅ are as defined for formula (I′),

to yield the compound of formula (VII):

wherein A₁, A₂, X, Y, R_(a), R_(b), R_(c), R_(d), T and R₅ are asdefined for formula (I′) and Alk is as defined hereinbefore,

the ester function of which compound of formula (VII) is hydrolysed toyield the corresponding carboxylic acid or carboxylate, which may beconverted into an acid derivative such as the corresponding acylchloride or anhydride before being coupled with an amine NHR₃R₄ whereinR₃ and R₄ have the same meanings as for formula (I′), before beingsubjected to the action of a pyrophosphate, phosphonate or phosphorylcompound under basic conditions, it being possible for the compoundthereby obtained to be optionally hydrolysed or hydrogenolysed to yieldthe compound of formula (I),

which compound of formula (I) may be purified according to aconventional separation technique, which is converted, if desired, intoits addition salts with a pharmaceutically acceptable acid or base andwhich is optionally separated into its isomers according to aconventional separation technique,

it being understood that, at any time considered appropriate in thecourse of the above-described process, certain groups (hydroxy, amino .. . ) of the reagents or intermediates of synthesis may be protected andthen deprotected according to the requirements of synthesis.

The compounds of formulae (II), (III), (VI) and the amine NHR₃R₄ areeither commercially available or can be obtained by the person skilledin the art using conventional chemical reactions described in theliterature.

More specifically, the phosphate compounds of formula (I) according tothe invention will be useful in the treatment of chemo- orradio-resistant cancers and also in malignant haemopathies andsmall-cell lung cancer.

Among the cancer treatments envisaged there may be mentioned, withoutimplying any limitation, cancers of the bladder, brain, breast anduterus, chronic lymphoid leukaemias, colorectal cancer, cancers of theoesophagus and liver, lymphoblastic leukaemias, non-Hodgkin lymphomas,melanomas, malignant haemopathies, myelomas, ovarian cancer,non-small-cell lung cancer, prostate cancer and small-cell lung cancer.Among non-Hodgkin lymphomas, there may be mentioned more preferablyfollicular lymphomas, mantle cell lymphomas, diffuse large B-celllymphomas, small lymphocytic lymphomas and marginal zone B-celllymphomas.

The present invention relates also to pharmaceutical compositionscomprising at least one compound of formula (I) in combination with oneor more pharmaceutically acceptable excipients.

Among the pharmaceutical compositions according to the invention theremay be mentioned more especially those that are suitable for oral,parenteral, nasal, per- or trans-cutaneous, rectal, perlingual, ocularor respiratory administration, especially tablets or dragées, sublingualtablets, sachets, paquets, capsules, glossettes, lozenges,suppositories, creams, ointments, dermal gels, and drinkable orinjectable ampoules.

The dosage varies according to the sex, age and weight of the patient,the administration route, the nature of the therapeutic indication, orof any associated treatments, and ranges from 0.01 mg to 1 g per 24hours in one or more administrations.

Furthermore, the present invention relates also to the association of acompound of formula (I) with an anticancer agent selected from genotoxicagents, mitotic poisons, anti-metabolites, proteasome inhibitors, kinaseinhibitors and antibodies, and also to pharmaceutical compositionscomprising that type of association and their use in the manufacture ofmedicaments for use in the treatment of cancer.

The compounds of the invention may also be used in association withradiotherapy in the treatment of cancer.

The following Preparations and Examples illustrate the invention withoutlimiting it in any way.

Preparation 1:6-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-1,3-benzodioxole-5-carboxylicacid Step A: 1-Formyl-2-piperidine-carboxylic acid

To a solution of 40 g of a racemic mixture of 2-piperidine-carboxylicacid (0.310 mmol) in 300 mL of formic acid placed at 0° C. there areadded, dropwise, 200 mL (2.15 mmol) of acetic anhydride. The batch isthen stirred at ambient temperature overnight. Then, the reactionmixture is cooled to 0° C., hydrolysed by adding 250 mL of water, andstirred for half an hour at 0° C. before being concentrated to dryness.The oil thereby obtained is taken up in 200 mL of methanol and thenconcentrated to dryness. The title product is obtained in the form of anoil in a yield of 98%. It is used directly, without being otherwisepurified, in the next Step.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 13.0 (m, 1H OH); 8.0-8.05 (2s, 1Haldehyde); 4.9-4.5 (2d, 1H α to the N and COOH); 4.1-2.6 (m, 2H α to theN); 2.2-1.2 (m, 6H piperidine)

IR: ν: —OH: 2000-3000 cm⁻¹ acid; ν: >C═O 1703 cm⁻¹ wide band

Step B: Methyl 5,6,7,8-tetrahydro-1-indolizine-carboxylate

To a solution of 10 g of the carboxylic acid obtained in Step A (63.6mmol) in 65 mL of dichloroethane there are successively added 13.4 g oftosyl chloride (70.4 mmol), 11.5 mL of methyl 2-chloroacrylate (113.5mmol) and then, dropwise, 17.8 mL of N,N,N-triethylamine (127.2 mmol).The reaction mixture is then refluxed for 1 hour 30 minutes. It is thenplaced at ambient temperature, and there are then added 5 mL of methyl2-chloroacrylate (48.9 mmol) and, dropwise, 9 mL of N,N,N-triethylamine(64 mmol). The batch is refluxed overnight. The reaction mixture is thendiluted with methylene chloride, washed successively with 1M HClsolution, saturated aqueous NaHCO₃ solution and then with brine until aneutral pH is obtained. The organic phase is then dried over MgSO₄,filtered, concentrated to dryness and purified by chromatography oversilica gel (heptane/AcOEt gradient). The title product is obtained inthe form of an oil.

¹H NMR: δ (400 MHz; CDCl₃; 300° K): 6.55-6.40 (d, 2H,tetrahydroindolizine); 3.91 (t, 3H methyl ester); 3.78 (s, 3Htetrahydroindolizine); 3.08 (t, 2H, tetrahydroindolizine); 1.95-1.85 (m,4H, tetrahydroindolizine)

IR: ν: >C═O 1692 cm⁻¹ ester

Step C: Methyl3-(6-formyl-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydro-1-indolizine-carboxylate

To a solution of 6.4 g of the ester obtained in Step B (35.7 mmol) in 12mL of N,N-dimethylacetamide, there are successively added 12.3 g of6-bromo-1,3-benzodioxole-5-carbaldehyde (53.6 mmol) and 7 g of potassiumacetate (71.4 mmol), and then the batch is stirred under argon for 20minutes. There are then added 1.3 g of palladium catalystdichlorobis(triphenylphosphine)palladium(II) (PdCl₂(PPh₃)₂) (1.8 mmol).The reaction mixture is then heated at 130° C. for one hour beforeadding 139 μL of H₂O thereto. Heating is maintained at that sametemperature overnight. The mixture is allowed to return to ambienttemperature and it is then diluted with AcOEt. Animal charcoal (25 g perg of product) is added and the batch is stirred at ambient temperaturefor 1 hour and then filtered. The organic phase is then washed withwater, dried over magnesium sulphate and concentrated to dryness. Thecrude product thereby obtained is purified by chromatography over silicagel (heptane/AcOEt gradient). The title product is obtained in the formof an oil.

¹H NMR: δ: (400 MHz; dmso-d6; 353° K): 9.65 (s, 1H, H aldehyde);7.3-7.15 (2s, 2H, aromatic Hs); 6.45 (s, 1H tetrahydroindolizine); 6.20(s, 2H methylenedioxy); 3.70 (s, 3H methyl ester); 3.5-4.0 (m, 2Htetrahydroindolizine); 3.05 (m, 2H tetrahydroindolizine); 1.85 (m, 4Htetrahydroindolizine) IR: ν: >C═O 1695 cm⁻¹ ester; ν: >C═O 1674 cm⁻¹

Step D:6-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]-1,3-benzodioxole-5-carboxylicacid

A solution containing 3.37 g of the compound obtained in Step C (10.3mmol) in 9.3 mL of acetone and 8.8 mL (80.24 mmol) of 2-methyl-2-buteneis prepared and placed at 0° C. There are added, dropwise, 9.3 mL of anaqueous solution containing a mixture of 3.3 g of sodium chlorite(NaClO₂) (36.05 mmol) and 3.6 g of sodium dihydrogen phosphatemonohydrate (NaH₂PO₄) (25.75 mmol). The batch is then stirred at ambienttemperature for 7 hours. The reaction mixture is then concentrated inorder to remove the acetone. The solid then obtained is filtered off,washed with water and then dried at 40° C. in vacuo overnight. The titleproduct is obtained in the form of a solid, which is subsequently usedwithout being otherwise purified.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 12.10 (m, 1H, H carboxylic acid);7.40-6.88 (2s, 2H, aromatic Hs); 6.20 (s, 1H, H tetrahydroindolizine);6.18 (s, 2H, H methylenedioxy); 3.70 (s, 3H, methyl ester); 3.55 (t, 2Htetrahydroindolizine); 3.00 (t, 2H tetrahydroindolizine); 1.80 (m, 4H, Htetrahydroindolizine)

IR: ν: —OH: 3000-2000 cm⁻¹ acid; ν: >C═O 1686-1676 cm⁻¹ ester+acid;ν: >C═C< 1608 cm⁻¹

Preparation 2:2-[1-(Methoxycarbonyl)-5,6,7,8-tetrahydro-3-indolizinyl]benzoic acid

The procedure is in accordance with the protocol described inPreparation 1, replacing the 6-bromo-1,3-benzodioxole-5-carbaldehydeused in Step C by 2-bromo-benzaldehyde.

Preparation 3:6-[1-(Methoxycarbonyl)-3-indolizinyl]-1,3-benzodioxole-5-carboxylic acidStep A: 1-(Carboxymethyl)-1,2-dihydropyridinium bromide

To a solution of 16.2 mL of pyridine (200 mmol) in 120 mL of ethylacetate there are added, in portions, 27.8 g (200 mmoles) of bromoaceticacid. The batch is then stirred at ambient temperature overnight. Theprecipitate thereby obtained is filtered off and then washed with coldethyl acetate. After drying, the title product is obtained in the formof a powder which is used directly in the next Step.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 9.15 (d, 2H, aromatic Hspyridine); 8.7 (t, 1H, aromatic H); 8.25 (t, 2H, aromatic H); 5.65 (s,2H, H CH₂COOH)

IR: ν: C═O: 1732 cm⁻¹; —OH acid: 2800 cm⁻¹

Step B: Methyl 1-indolizinecarboxylate

To a suspension of 6.55 g of the pyridinium salt obtained in Step A (30mmol) in 240 mL of toluene there are successively added 16.7 mL ofmethyl acrylate (150 mmol), 4.2 mL of triethylamine (30 mmol) and then,in portions, 20.9 g of MnO₂ (240 mmol). The batch is then heated at 90°C. for 3 hours. After cooling, the reaction mixture is filtered over acake of Celite and concentrated to dryness. The title product is thenisolated by purification over silica gel (heptane/AcOEt gradient: 0-10%)in the form of an oil which crystallises in the cold state.

¹H NMR: δ (300 MHz; dmso-d6; 300° K): 8.5 (d, 1H, H indolizine); 8.05(d, 1H, H indolizine); 7.6 (s, 1H, H indolizine); 7.15 (m, 2H, Hindolizine); 6.85 (m, 1H, H indolizine); 4.25 (q, 2H, —C(O)CH₂CH₃); 1.35(t, 3H, —C(O)CH₂CH₃)

IR: ν: C═O ester: 1675 cm⁻¹; aromatic C═C moieties: 1634 cm⁻¹

Step C:6-[1-(Methoxycarbonyl)-3-indolizinyl]-1,3-benzodioxole-5-carboxylic acid

The procedure is in accordance with the protocol described in Steps Cand D of Preparation 1.

Preparation 4:4-Chloro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]-benzoic acidStep A: Ethyl 1,2-dimethyl-1H-pyrrole-3-carboxylate

To a solution of 10 g of ethyl 2-methyl-1H-pyrrole-3-carboxylate (65.3mmol) and 8.95 mL (130.6 mmol) of methyl iodide in 70 mL ofdimethylformamide placed at 0° C. there are added, in three portions,2.61 g (65.3 mmol) of sodium hydride 60%. The batch is then stirred at0° C. for 1 hour. Then, the reaction mixture is hydrolysed by theaddition of 420 mL of ice-cold water. The reaction mixture is thendiluted with ethyl acetate, successively washed with 0.1M HCl solution,saturated aqueous LiCl solution and then brine. The organic phase isthen dried over MgSO₄, filtered, concentrated to dryness and purified bychromatography over silica gel (petroleum ether/AcOEt gradient).

¹H NMR: δ (400 MHz; dmso-d6; 300K): 6.65 (d, 1H pyrrole); 6.3 (1d, 1Hpyrrole); 4.1 (1q, 2H, OCH₂CH₃); 3.5 (s, 3H N-pyrrole); 2.4 (s, 3Hpyrrole); 1.5 (1t, 3H OCH₂CH₃)

IR: ν: >C═O: 1688 cm⁻¹; ν: C—O—C: 1172 cm⁻¹

Step B: Ethyl5-(5-chloro-2-formylphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxylate

To a solution of 10.5 g of the compound obtained in Step A (62.8 mmol)in 65 mL of N,N-dimethylacetamide there are successively added 15.2 g of2-bromo-4-chlorobenzaldehyde (69 mmol), 12.3 g of potassium acetate(125.6 mmol) and then the batch is stirred under argon for 20 minutes.There are then added 2.2 g of palladium catalyst PdCl₂(PPh₃)₂ (3.14mmol). The reaction mixture is then heated at 130° C. overnight. Themixture is allowed to return to ambient temperature and it is thendiluted with dichloromethane. Animal charcoal is added (30 g) and thebatch is stirred at ambient temperature for 1 hour and then filtered.The organic phase is then washed with water, dried over magnesiumsulphate and concentrated to dryness. The crude product thereby obtainedis purified by chromatography over silica gel (petroleum ether/AcOEtgradient). The title product is obtained in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 9.8 (s, 1H, formyl); 7.91-7.69-7.61(d, 3H, aromatic Hs); 6.5 (s, 1H pyrrole); 4.2 (q, 2H, OCH₂CH₃); 3.4 (s,3H, CH₃—N-pyrrole); 2.55 (s, 3H pyrrole); 1.28 (t, 3H, OCH₂CH₃)

Step C:4-Chloro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benzoic acid

A solution is prepared containing 12.85 g of the compound obtained inStep B (42 mmol) and 35.7 mL (336 mmol) of 2-methyl-2-butene in amixture containing 20 mL of acetone and 20 mL of tetrahydrofuran. Thereare added, dropwise, 200 mL of an aqueous solution containing a mixtureof 13.3 g of sodium chlorite (NaClO₂) (147 mmol) and 14.5 g of sodiumdihydrogen phosphate monohydrate (NaH₂PO₄H₂O) (105 mmol). The batch isthen vigorously stirred at ambient temperature for 7 hours. The reactionmixture is then concentrated to remove the acetone. Ethyl acetate isadded, and the organic phase is washed with water and then concentratedto dryness. The residue is then taken up in a minimum of ethyl ether.The solid then obtained is filtered off, washed with ether and thendried in vacuo at 40° C. overnight. The title product is obtained in theform of a solid, which is subsequently used without being otherwisepurified.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 13 (m, 1H COOH); 7.85-7.6-7.41(d,dd,wd, 3H, aromatic Hs); 6.3 (s, 1H, H pyrrole); 4.15 (q, 2H, OCH₂CH₃);3.25 (s, 3H, CH₃—N-pyrrole); 2.5 (s, 3H, CH₃-pyrrole); 1.25 (t, 3H,OCH₂CH₃)

IR: ν: —OH: 3100-2500 cm⁻¹ acid; ν: >C═O: 1681 cm⁻¹ ester+acid

Preparation 5:6-[4-(Ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]-1,3-benzodioxole-5-carboxylicacid

The procedure is in accordance with the process of Preparation 4,replacing the 2-bromo-4-chlorobenzaldehyde used in Step B by6-bromo-1,3-benzodioxole-5-carbaldehyde.

Preparation 6:4-Fluoro-3-methoxy-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benzoicacid

The procedure is in accordance with the process of Preparation 4,replacing the 2-bromo-4-chlorobenzaldehyde used in Step B by2-bromo-4-fluoro-3-methoxybenzaldehyde.

Preparation 7:4-Fluoro-2-[4-(ethoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]benzoic acid

The procedure is in accordance with the process of Preparation 4,replacing the 2-bromo-4-chlorobenzaldehyde used in Step B by2-bromo-4-fluorobenzaldehyde.

Preparation 8:7-[4-(Methoxycarbonyl)-1,5-dimethyl-1H-pyrrol-2-yl]-2,3-dihydro-1,4-benzodioxin-6-carboxylicacid

The procedure is in accordance with the process of Preparation 4,replacing the ethyl 2-methyl-1H-pyrrole-3-carboxylate in Step A bymethyl 2-methyl-1H-pyrrole-3-carboxylate and the2-bromo-4-chlorobenzaldehyde used in Step B by7-bromo-2,3-dihydro-1,4-benzodioxin-6-carbaldehyde.

Preparation 9:5-Benzyloxy-2-(1-methoxycarbonyl-5,6,7,8-tetrahydroindolizin-3-yl)benzoicacid Step A: Methyl3-(4-benzyloxy-2-formyl-phenyl)-5,6,7,8-tetrahydroindolizine-1-carboxylate

5-Benzyloxy-2-bromo-benzaldehyde (12.3 g, 42.2 mmol) is introduced intoa flask in the presence of potassium acetate (8.3 g; 84.2 mmol) and 120mL of dimethylacetamide. After degassing under argon,dichlorobis(triphenylphosphine)palladium (II) (1.04 g, 1.5 mmol) isadded and the mixture is then degassed under argon before being heatedat 100° C. for 16 hours. After returning to ambient temperature, thereaction mixture is poured into 200 mL of ethyl acetate, filtered overCelite, and washed with water and then with brine. The combined aqueousphases are extracted with ethyl acetate. The organic phases are driedover sodium sulphate, filtered and concentrated under reduced pressure.The residue obtained is purified by chromatography over silica gel inorder to obtain the title product.

Step B:5-Benzyloxy-2-(1-methoxycarbonyl-5,6,7,8-tetrahydroindolizin-3-yl)benzoicacid

To a solution of the compound obtained in Step B (4.63 g, 11.89 mmol) in300 mL of acetone there is added 2-methyl-2-butene (6.31 mL, 59 mmol). Asolution of sodium dihydrogen phosphate monohydrate (6.56 g, 47.6 mmol)and sodium chlorite (2.69 g, 23.8 mmol) in 40 mL of water is then pouredin dropwise whilst maintaining the temperature below 20° C. Afterstirring for 30 minutes at ambient temperature, the mixture is acidifiedwith 2M HCl solution and then the phases are separated. The aqueousphase is extracted with ethyl acetate. The combined organic phases aredried over sodium sulphate, filtered and evaporated to dryness toprovide the expected compound.

Preparation 1′:(3S)-3-(4-Morpholinylmethyl)-1,2,3,4-tetrahydroisoquinoline Step A:Benzyl(3S)-3-(4-morpholinylcarbonyl)-3,4-dihydro-2(1H)-isoquinoline-carboxylate

To a solution of 5 g of(3S)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydro-3-isoquinolinecarboxylicacid (16 mmol) in 160 mL of dichloromethane there are added 1.5 mL ofmorpholine (17.6 mmol), then 9 mL of N,N,N-triethylamine (64 mmol), 3.3g of 1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (EDC) (19.2 mmol)and 2.6 g of hydroxybenzotriazole (HOBt) (19.2 mmol). The reactionmixture is stirred at ambient temperature overnight; it is then pouredinto aqueous ammonium chloride solution and extracted with ethylacetate. The organic phase is then dried over magnesium sulphate, andthen filtered and evaporated to dryness. The crude product therebyobtained is then purified by chromatography over silica gel(dichloromethane/methanol gradient). The product is obtained in the formof a foam.

¹H NMR: δ (400 MHz; dmso-d6; 353K): 7.30 (m, 5H benzyl); 7.15 (m, 4H,aromatic Hs); 5.2-5.0 (m, 3H, 2H benzyl, 1H dihydroisoquinoline);4.75-4.5 (2d, 2H dihydroisoquinoline); 3.55-3.3 (m, 8H morpholine);3.15-2.9 (2dd, 2H dihydroisoquinoline)

IR: ν: >C═O: 1694; 1650 cm⁻¹

Step B: Benzyl(3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinecarboxylate

To a solution of 5.3 g of the product obtained in Step A (13.9 mmol) in278 mL of tetrahydrofuran there are added 14 mL ofborane-dimethylsulphide complex (BH₃Me₂S) (27.8 mmol) at ambienttemperature. The batch is heated for 4 hours at 80° C. It is allowed toreturn to ambient temperature and there are then added 7 mL (14 mmol) ofBH₃Me₂S. The reaction mixture is again heated at 80° C. for 2 hours. Thetetrahydrofuran is then evaporated off and then there is slowly addedmethanol and then 5.6 mL of 5M hydrochloric acid (27.8 mmol). Themixture is stirred at ambient temperature overnight, and then at 80° C.for 1 hour. Saturated aqueous NaHCO₃ solution is then added to thereaction mixture placed at 0° C. until a pH of 8 is obtained, andextraction with ethyl acetate is then carried out. The organic phase isthen dried over magnesium sulphate, and then filtered and evaporated todryness. The title product is obtained in the form of an oil.

¹H NMR: δ (400 MHz; dmso-d6; 353K): 7.43-7.30 (unresolved peak, 5Hbenzyl); 7.19 (m, 4H, aromatic Hs); 5.16 (m, 2H, 2H benzyl); 4.79-4.29(d, 2H dihydroisoquinoline); 4.58 (m, 1H dihydroisoquinoline); 3.50 (m,4H morpholine); 3.02-2.80 (dd, 2H dihydroisoquinoline); 2.42-2.28(unresolved peak, 5H, 4H morpholine, 1H morpholine); 2.15 (dd, 1Hmorpholine)

IR: ν: >CH: 2810 cm⁻¹; ν: >C═O: 1694 cm⁻¹; ν: >C—O—C<: 1114 cm⁻¹;ν: >CH—Ar: 751; 697 cm⁻¹

Step C: (3S)-3-(4-Morpholinylmethyl)-1,2,3,4-tetrahydroisoquinoline

To a solution of 4.9 g of the compound of Step B (13.4 mmol) in 67 mL ofethanol there is added 0.980 g of palladium dihydroxide (20% by weight)at ambient temperature. The reaction mixture is placed under 1.2 bars ofhydrogen at ambient temperature for 4 hours. It is then passed through aWhatman filter and the palladium is then rinsed several times withethanol. The filtrate is evaporated to dryness. The title product isobtained in the form of an oil.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.12-7.0 (unresolved peak, 4H,aromatic Hs); 3.92 (s, 2H tetrahydroisoquinoline); 3.60 (t, 4Hmorpholine); 2.98 (m, 1H tetrahydroisoquinoline); 2.68 (dd, 1Htetrahydroisoquinoline); 2.5-2.3 (unresolved peak, 8H, 1Htetrahydroisoquinoline, 6H morpholine, 1H NH)

IR: ν: >NH: 3322 cm^(−1;) ν: >C—O—C<: 1115 cm^(−1;) ν: >CH—Ar: 742 cm⁻¹

Preparation 2′: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinolinehydrochloride Step A:{(3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl4-methylbenzenesulphonate

To a solution of 30.2 g of[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methanol (185 mmol) in 750 mLof dichloromethane there are successively added 91.71 g of tosylchloride (481 mmol) and then, dropwise, 122.3 mL of N,N,N-triethylamine(740 mmol). The reaction mixture is then stirred at ambient temperaturefor 20 hours. It is then diluted with dichloromethane, washedsuccessively with 1M HCl solution, saturated aqueous NaHCO₃ solution andthen brine until neutral. The organic phase is then dried over MgSO₄,filtered and concentrated to dryness. The solid obtained is thendissolved in a minimum volume of dichloromethane and then cyclohexane isadded until a precipitate is formed. This precipitate is then filteredoff and washed with cyclohexane. After drying, the title product isobtained in the form of crystals.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 7.75 (d, 2H, aromatic Hs, orthoO-tosyl); 7.6 (d, 2H, aromatic Hs, ortho N-tosyl); 7.5 (d, 2H, aromaticHs, meta O-tosyl); 7.3 (d, 2H, aromatic Hs, meta N-tosyl); 7.15-6.9 (m,4H, aromatic Hs, tetrahydroisoquinoline); 4.4-4.15 (dd, 2H, aliphaticHs, tetrahydroisoquinoline); 4.25 (m, 1H, aliphatic H,tetrahydroisoquinoline); 4.0-3.8 (2dd, 2H, aliphatic Hs, CH₂—O-tosyl);2.7 (2dd, 2H, aliphatic Hs, tetrahydroisoquinoline); 2.45 (s, 3H,O—SO₂-Ph-CH₃); 2.35 (s, 3H, N—SO₂-Ph-CH₃)

IR: ν: —SO₂: 1339-1165 cm⁻¹

Step B:(3R)-3-Methyl-2-[(4-methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinoline

To a suspension of 8.15 g (214.8 mmol) of LiAlH₄ in 800 mL of methyltert-butyl ether (MTBE) there are added 101.2 g of the ditosyl compoundobtained in Step A (214.8 mmol) dissolved in 200 mL of MTBE. The batchis then heated at 50° C. for 2 hours. It is allowed to cool and placedat 0° C., and there are then added, dropwise, 12 mL of 5M NaOH solution.The batch is stirred at ambient temperature for 45 minutes. The solidthereby obtained is then filtered off and washed with MTBE and then withdichloromethane. The filtrate is then concentrated to dryness. The titleproduct is then obtained in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 7.70 (d, 2H, aromatic Hs, orthoN-tosyl); 7.38 (d, 2H, aromatic Hs, meta N-tosyl); 7.2-7.0 (m, 4H,aromatic Hs, tetrahydroisoquinoline); 4.4 (m, 2H, aliphatic Hs,tetrahydroisoquinoline); 4.3 (m, 1H, aliphatic H,tetrahydroisoquinoline); 2.85-2.51 (2dd, 2H, aliphatic Hs,tetrahydroisoquinoline); 2.35 (s, 3H, N—SO₂-Ph-CH₃); 0.90 (d, 3H,tetrahydroisoquinoline-CH₃)

IR: ν: —SO₂: 1332-1154 cm⁻¹

Step C: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline

To a solution of 31.15 g (103.15 mmol) of the monotosyl compoundobtained in Step B in 500 mL of anhydrous methanol there are added, inportions, 3.92 g (161 mmol) of magnesium turnings. The batch is stirredin the presence of ultrasound for 96 hours. The reaction mixture is thenfiltered and the solid is washed several times with methanol. Thefiltrate is then concentrated to dryness. After purification bychromatography over silica gel (dichloromethane/EtOH/NH₄OH gradient),the title product is obtained in the form of an oil.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 7.05 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 3.90 (m, 2H, aliphatic Hs,tetrahydroisoquinoline); 2.85 (m, 1H, aliphatic H,tetrahydroisoquinoline); 2.68-2.4 (2dd, 2H, aliphatic Hs,tetrahydroisoquinoline); 1.12 (d, 3H, tetrahydroisoquinoline-CH₃);2.9-2.3 (m, broad, 1H, HN (tetrahydroisoquinoline))

IR: ν: —NH: 3248 cm⁻¹

Step D: (3R)-3-Methyl-1,2,3,4-tetrahydroisoquinoline hydrochloride

To a solution of 14.3 g (97.20 mmol) of the compound obtained in Step Cin 20 mL of anhydrous ethanol there are added, dropwise, 100 mL of a 1Msolution of HCl in ether. The batch is stirred at ambient temperaturefor 1 hour and then filtered. The crystals thereby obtained are washedwith ethyl ether. After drying, the title product is obtained in theform of crystals.

¹H NMR: δ (400 MHz; dmso-d6; 300° K): 9.57 (m, broad, 2H, NH₂ ⁺(tetrahydroisoquinoline); 7.22 (m, 4H, aromatic Hs,tetrahydroisoquinoline); 4.27 (s, 2H, aliphatic Hs,tetrahydroisoquinoline); 3.52 (m, 1H, aliphatic H,tetrahydroisoquinoline); 3.03-2.85 (2dd, 2H, aliphatic Hs,tetrahydroisoquinoline); 1.39 (d, 3H, tetrahydroisoquinoline-CH₃)

IR: ν: —NH₂ ⁺: 3000-2300 cm⁻¹; ν: aromatic —CH: 766 cm⁻¹

Preparation 3′:(3R)-3-[3-(Morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline Step A:{(3S)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl4-methylbenzenesulphonate

The procedure is the same as that of Step A of Preparation 2′.

Step B: tert-Butyl2-({(3R)-2-[(4-methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl)-3-(morpholin-4-yl)-3-oxopropanoate

To a suspension of 1 g of NaH (60%) (25.08 mmol) in 30 mL of MTBE thereare added, dropwise, a solution of 5 g of tert-butyl3-morpholino-3-oxopropanoate (21.81 mmol) in 20 mL of anhydrous MTBE.This suspension is stirred at ambient temperature for 1 hour and thenthe compound obtained in Step A is added in the form of a powder. Thebatch is stirred at 60° C. for 30 hours. 100 mL of saturated aqueousammonium chloride solution are added. The resulting solution isextracted with dichloromethane. The organic phase is then dried overMgSO₄, filtered and concentrated to dryness. After purification bychromatography over silica gel (dichloromethane/MeOH gradient), theexpected product is obtained in the form of an oil.

¹H NMR (500 MHz, dmso-d6) δ ppm: 7.63/7.59 (2d, 2H), 7.3/7.26 (2d, 2H),7.13 (m, 2H), 7.09/6.97 (2t, 2H), 4.64/4.55/4.36/4.28 (2AB, 2H),4.25/4.11 (2m, 1H), 3.81 (m, 1H), 3.73-3.48 (m, 4H), 3.57-3.32 (m, 4H),2.51 (m, 2H), 2.32/2.31 (2s, 3H), 1.88/1.79 (2m, 2H), 1.39/1.38 (2s, 9H)

IR (ATR) cm⁻¹: ν: >C═O: 1731 (ester); ν: >C═O: 1644 (amide); ν: —SO2:1334-1156; ν: >C—O—C<: 1115; γ: >CH—Ar: 815-746-709

Step C:2-({(3R)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}methyl)-3-(morpholin-4-yl)-3-oxopropanoicacid

To a solution of 9.5 g (17.97 mmol) of the compound obtained in Step Bin 40 mL of dioxane there are added, dropwise, 20 mL of a 4M solution ofHCl in dioxane. The batch is stirred at ambient temperature for 48 hoursand then the solution is concentrated to dryness. After drying, theexpected product is obtained in the form of an oil.

¹H NMR (400 MHz, dmso-d6) δ ppm: 12.75 (m, 1H), 7.6 (2*d, 2H), 7.3 (2*d,2H), 7.1/6.95 (2*m, 4H), 4.7-4.2 (d, 2H), 4.25/4.12 (2*m, 1H), 3.9-3.3(m, 9H), 2.55 (d, 2H), 2.3 (2*s, 3H), 1.8 (t, 2H)

IR (ATR) cm⁻¹: ν: —OH : 3500 to 2000; ν: >C═O: 1727 (acid); ν: >C═O:1634 (amide); ν: —SO2: 1330-1155

Step D:3-{(3R)-2-[(4-Methylphenyl)sulphonyl]-1,2,3,4-tetrahydroisoquinolin-3-yl}-1-(morpholin-4-yl)propan-1-one

To a solution of 7.80 g (16.51 mmol) of the compound obtained in Step Cin 100 mL of DMSO there are added 1.16 g (19.83 mmol) of solid sodiumchloride and then, dropwise, 5 mL of water. The batch is stirred at 130°C. for 1 hour and then the solution is concentrated to ¾. The reactionmixture is then diluted with dichloromethane and washed successivelywith saturated aqueous lithium chloride solution and then with brine.The organic phase is then dried over MgSO₄, filtered and concentrated todryness. After purification by chromatography over silica gel(cyclohexane/ethyl acetate gradient), the expected product is obtainedin the form of an oil.

¹H NMR (400 MHz, dmso-d6) δ ppm: 7.65 (d, 2H), 7.3 (d, 2H), 7.15/7 (2 m,4H), 4.6 (d, 1H), 4.25 (d, 1H), 4.2 (m, 1H), 3.5 (m, 4H), 3.4 (2 m, 4H),2.6 (2 dd, 2H), 2.35 (s, 3H), 2.3 (m, 2H), 1.5 (quad., 2H)

IR (ATR) cm⁻¹: ν: >C═O: 1639; ν: —SO2: 1331-1156; γ: >CH—Ar: 815-675

Step E:(3R)-2-[(4-Methylphenyl)sulphonyl]-3-[3-(morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline

To a solution of 6.0 g (14.0 mmol) of the compound obtained in Step D in60 mL of MTBE and 14 mL of dichloromethane there are added 1.06 g (28mmol) of LAH in portions over 5 minutes. The batch is stirred at ambienttemperature for 15 hours. There are added, dropwise, 1.5 mL of water andstirring is carried out for 15 minutes. There are then added, dropwise,1.5 mL of 5M sodium hydroxide solution and stirring is carried out for15 minutes. The reaction mixture is then diluted with MTBE anddichloromethane. The suspension is then filtered and the precipitate iswashed with MTBE and dichloromethane. The organic phase is then driedover MgSO₄, filtered and concentrated to dryness. After purification bychromatography over silica gel (dichloromethane/EtOH/NH₄OH gradient),the expected product is obtained in the form of an oil.

¹H NMR (400 MHz, dmso-d6) δ ppm: 7.68 (d, 2H), 7.32 (d, 2H), 7.1(unresolved peak, 4H), 4.65/4.23 (AB, 2H), 4.2 (m, 1H), 3.55 (t, 4H),2.7/2.6 (ABx, 2H), 2.35 (s, 3H), 2.25 (t, 4H), 2.2 (t, 2H), 1.4/1.3 (2m,4H).

IR (ATR) cm⁻¹: ν: —SO2: 1333-1158

Step F: (3R)-3-[3-(Morpholin-4-yl)propyl]-1,2,3,4-tetrahydroisoquinoline

To a solution of 1.50 g (3.62 mmol) of the compound obtained in Step Ein 20 mL of anhydrous methanol there are added 2.0 g (82.3 mmol), inportions, of magnesium turnings. The batch is stirred in the presence ofultrasound for 96 hours. The reaction mixture is then filtered, thesolid is washed several times with methanol, and the filtrate isconcentrated to dryness. After purification by chromatography oversilica gel (dichloromethane/EtOH/NH₄OH gradient), the expected productis obtained in the form of an oil.

¹H NMR (400 MHz, dmso-d6) δ ppm: 7.3 (d, 2H), 7.1 (t, 2H), 7.1 (d+t,3H), 7 (d, 2H), 3.9 (s, 2H), 3.55 (t, 4H), 2.75 (m, 1H), 2.72/2.45 (dd,2H), 2.35 (t, 4H), 2.25 (t, 2H), 1.6 (m, 2H), 1.45 (m, 2H)

IR (ATR) cm⁻¹: ν: >NH2+/NH+: 3500-2300; ν: >C—O—C<: 1115

High-Resolution Mass Spectrometry (ESI+/FIA/HR):

Empirical formula: C₁₆H₂₄N₂O

[M+H]⁺ calculated: 261.1961

[M+H]⁺ measured: 261.1959

Preparation 4′:(3R)-3-(4-Morpholinylmethyl)-1,2,34-tetrahydroisoquinoline

The procedure is in accordance with the process of Preparation 1′,replacing the(3S)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydro-3-isoquinolinecarboxylicacid used in Step A by(3R)-2-[(benzyloxy)carbonyl]-1,2,3,4-tetrahydro-3-isoquinolinecarboxylicacid.

Preparation 1″: 4-{[tert-Butyl(dimethyl)silyl]oxy}-N-phenylaniline

To a solution of 12 g of 4-anilinophenol (64.7 mmol) in 200 mL ofacetonitrile there are added, at ambient temperature, 6.7 g of imidazole(97.05 mmol) and 11.7 g of tert-butyl(chloro)dimethylsilane (77.64mmol). The batch is stirred at 70° C. for 4 hours. The reaction mixtureis then poured into water and extracted with ether. The organic phase isthen dried over magnesium sulphate, then filtered and evaporated todryness. The crude product thereby obtained is then purified bychromatography over silica gel (petroleum ether/dichloromethanegradient). The title product is obtained in the form of a powder.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.84 (s, 1H NH); 7.17 (t, 2Haniline); 6.98 (d, 2H phenoxy); 6.94 (d, 2H aniline); 6.76 (d, 2Hphenoxy); 6.72 (t, 1H aniline); 0.95 (s, 9H tert-butyl); 0.15 (s, 6Hdimethyl)

IR: ν: >NH: 3403 cm⁻¹; ν: >Ar: 1597 cm⁻¹

Preparation 2″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-indol-5-amine

The procedure is in accordance with the process of Preparation 5″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B by5-bromo-1-methyl-1H-indole.

Preparation 3″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-pyrrolo[2,3-b]pyridin-5-amine

The procedure is in accordance with the process of Preparation 5″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B by5-bromo-1-methyl-1H-pyrrolo[2,3-b]pyridine (obtained in accordance witha protocol from the literature: Heterocycles, 60(4), 865, 2003).

IR: ν: —NH—: 3278 cm⁻¹; ν: aromatic —C═C— moieties: 1605 cm⁻¹

Preparation 4″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)pyridin-4-amine

The procedure is in accordance with the process of Preparation 5″,replacing the 4-bromo-1-methyl-1H-pyrazole used in Step B by4-bromopyridine.

IR: ν —NH—: 3200 and 2500 cm⁻¹; ν —Si—O—: 902 cm⁻¹; ν —Si—C—: 820 cm⁻¹

Preparation 5″:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-1-methyl-1H-pyrazol-4-amineStep A: 4-{[tert-Butyl(dimethyl)silyl]oxy}aniline

The title compound is obtained starting from 4-aminophenol in THF in thepresence of imidazole and tert-butyl(chloro)dimethylsilane in accordancewith the protocol described in the literature (S. Knaggs et al, Organic& Biomolecular Chemistry, 3(21), 4002-4010; 2005).

¹H NMR: δ (400 MHz; dmso-d6; 300K): 6.45-6.55 (dd, 4H, aromatic Hs);4.60 (m, 2H, NH₂-Ph); 0.90 (s, 9H, Si (CH₂)₂CH(CH₃)₂); 0.10 (s, 6H, Si(CH₂)₂CH(CH₃)₂)

IR: ν: —NH₂ ⁺: 3300-3400 cm⁻¹

Step B:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-1-methyl-pyrazol-4-amine

To a solution of 30.8 g (0.137 mol) of the compound of Step A in 525 mLof anhydrous toluene there are successively added 29.8 g of sodiumtert-butylate (0.310 mol), 4.55 g of Pd₂(dba)₃ (also referred to astris(dibenzylideneacetone)dipalladium(0)) (4.96 mmol), 4.81 g of2-di-tert-butylphosphino-2′,4′,6′-tri-isopropyl-1,1′-biphenyl (9.91mmol) and 12.8 mL of 4-bromo-1-methyl-1H-pyrazole (0.124 mol). The batchis degassed under argon for 30 minutes and then refluxed for 3 hours. Itis allowed to cool. The reaction mixture is concentrated to dryness andthen taken up in dichloromethane, filtered over Celite and thenconcentrated to dryness again. The residue is then purified bychromatography over silica gel (gradient CH₂Cl₂/AcOEt) to provide theexpected product in the form of a solid.

¹H NMR: δ (400 MHz; dmso-d6; 300K): 7.55 (s, 1H, pyrazole); 7.23 (s, 1H,pyrazole); 7.18 (broad s, 1H, NH₂-Ph); 6.64 (m, 4H, aromatic Hs); 3.77(s, 3H, CH₃-pyrazole); 0.90 (s, 9H, Si (CH₂)₂CH(CH₃)₂); 0.12 (s, 6H, Si(CH₂)₂CH(CH₃)₂)

IR: ν —NH⁺: 3275 cm⁻¹; ν Ar and C═N: 1577 and 1502 cm⁻¹; ν —Si—C—: 1236cm⁻¹; ν —Si—O—: 898 cm⁻¹; ν —Si—C—: 828, 774 cm⁻¹

Preparation 6″:N-{4-[(tert-Butyldimethylsilyl)oxy]phenyl}-1-trideuteriomethyl-1H-pyrazol-4-amineStep A: 4-Bromo-1-trideuteriomethyl-1H-pyrazole

4-Bromo-1H-pyrazole (9.05 g, 61.6 mmol) is added in portions to asuspension of sodium hydride (60% in oil) (2.83 g, 70.8 mmol) intetrahydrofuran (90 mL) cooled in an ice bath. After having taken awaythe ice bath, the solution is stirred at ambient temperature for 0.5hours. It is again cooled in an ice bath and iodomethane-d₃ (5.0 mL,80.3 mmol) is added. The solution is stirred at ambient temperature for19 hours. The suspension is then concentrated. The evaporation residueis triturated with tert-butyl methyl ether (90 mL) and filtered. Thefiltrate is concentrated in vacuo to obtain the expected compound in theform of an oil.

¹H NMR (400 MHz, CDCl₃) δ ppm: 7.37 (s, 1H), 7.43 (s, 1H)

Step B:N-{4-[(tert-Butyldimethylsilyl)oxy]phenyl}-1-trideuteriomethyl-1H-pyrazol-4-amine

4-Bromo-1-trideuteriomethyl-1H-pyrazole (9.6 g, 58.5 mmol),4-[(tert-butyldimethyl-silyl)oxy]aniline (14.4 g, 64.6 mmol) and toluene(150 mL) are added to a 500-ml three-necked flask. The solution isdegassed with nitrogen for 15 minutes, and then sodium tert-butylate(11.4 g, 0.12 mol),2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl (0.77 g, 1.81mmol) and tris(dibenzylideneacetone)dipalladium(0) (1.64 g, 1.79 mmol)are successively added. The suspension is heated at 85° C. for 1.5hours. The reaction mixture is then cooled to ambient temperature andwater (270 mL) is added. The mixture is stirred for 30 minutes. Celite(30 g) is then added and the suspension is filtered on a bed of Celite.The phases of the filtrate are separated and the aqueous phase isextracted with ethyl acetate (3×200 mL). The combined organic phases aredried over sodium sulphate and filtered. The product is purified bychromatography over silica gel (ethyl acetate/heptane gradient). Theproduct obtained is recrystallized from heptane (80 mL) to obtain theexpected compound.

¹H NMR (400 MHz, CDCl₃) δ ppm: 0.16 (s, 6H), 0.97 (s, 9H), 4.92 (s, 1H),6.61-6.73 (m, 4H), 7.25 (s, 1H), 7.36 (s, 1H)

¹³C NMR (100 MHz, CDCl₃) δ ppm: −4.37, 18.28, 25.86, 38.67 (sept.,¹J_(C-D)=21.0 Hz), 115.12, 120.73, 123.76, 126.52, 134.74, 141.07,148.43

MS (ESI): [M+H]⁺ 307.08

Preparation 7″:4-({4-[(tert-Butyldimethylsilyl)oxy]phenyl}amino)-1,5-dimethyl-1H-pyrrole-2-carbonitrileStep A: 4-Bromo-1,5-dimethyl-1H-pyrrole-2-carbonitrile

A solution of bromine (6.58 mL, 0.13 mol) in acetic acid (60 mL) isadded dropwise, with the aid of a dropping funnel, to a solution of1,5-dimethyl-1H-pyrrole-2-carbonitrile (15.0 g, 0.12 mol) in acetic acid(300 mL). The batch is stirred at ambient temperature for 24 hours. Thereaction mixture is then poured into a beaker containing 300 mL ofwater. The solid formed is filtered off and rinsed with water. It isthen dissolved in dichloromethane (300 mL) and the organic phase iswashed with brine, dried over sodium sulphate, filtered and concentratedin vacuo to yield the expected product in the form of a solid.

¹H NMR (CDCl₃) δ ppm: 2.25 (s, 3H), 3.67 (s, 3H), 6.74 (s, 1H)

Step B:4-({4-[(tert-Butyldimethylsilyl)oxy]phenyl}amino)-1,5-dimethyl-1H-pyrrole-2-carbonitrile

A solution of the compound of the above Step (1.5 g, 7.53 mmol),4-[(tert-butyldimethylsilyl)oxy]aniline (2.02 g, 9.04 mmol), sodiumtert-butylate (1.45 g, 15.06 mmol) and2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl (0.13 g, 0.30mmol) in toluene (20 mL) is purged with nitrogen.Tris(dibenzylideneacetone)-dipalladium(0) (0.28 g, 0.30 mmol) is added,and then the reaction mixture is heated at 90° C. until the reaction iscomplete (monitored by TLC). Heating is stopped and the mixture isallowed to return to ambient temperature. Water (75 mL) is added and themixture is extracted with ethyl acetate (3×75 mL). The combined organicphases are washed with brine and then concentrated. The crude product ispurified by flash chromatography over silica gel (ethyl acetate/heptanegradient). The product thereby obtained is dissolved in heptane in thewarm state and is allowed to precipitate, with stirring, at ambienttemperature, and then at 0° C. The solid is filtered off and theoperation is repeated on the filtrate to yield the expected compound inthe form of a solid.

¹H NMR (400 MHz, CDCl₃) δ ppm: 0.15 (s, 6H), 0.97 (s, 9H), 2.13 (s, 3H),3.66 (s, 3H), 4.68 (br. s, 1H), 6.49 (d, J=8.5 Hz, 2H), 6.64 (s, 1H),6.66 (d, J=8.7 Hz, 2H)

¹³C NMR (100 MHz, CDCl₃) δ ppm: 4.34, 9.72, 18.30, 25.88, 32.94, 101.27,114.37, 114.70, 116.41, 120.73, 124.52, 131.23, 141.54, 148.27

MS (ESI+): [M+H]⁺ measured: 342.3

Preparation 8″:4-[(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)amino]-1-methyl-1H-pyrrole-2-carbonitrileStep A: 1-Methyl-1H-pyrrole-2-carbonitrile

N,N-Dimethylformamide (3 mL) and 1,4-diazabicyclo[2.2.2]octane (0.49 g,4.3 mmol) are added to a solution of pyrrole-2-carbonitrile (4 g, 43.4mmol) in dimethyl carbonate (56 mL). The solution is stirred at 90° C.for 15 hours, and is then heated at 110° C. for 8 hours. The mixture iscooled to ambient temperature, and then ethyl acetate (80 mL) is added.The phases are separated and the organic phase is washed with water(2×80 mL) and 1M aqueous hydrochloric acid solution (1×80 mL). Thecombined aqueous phases are extracted again with ethyl acetate (1×80mL). The combined organic phases are washed with brine (1×80 mL), driedover magnesium sulphate, filtered and concentrated in vacuo to obtainthe expected product in the form of a liquid.

¹H NMR (400 MHz, CDCl₃) δ ppm: 3.78 (m, 2H), 6.12-6.18 (m, 1H),6.74-6.82 (m, 1H)

Step B: 4-Bromo-1-methyl-1H-pyrrole-2-carbonitrile

N-Bromosuccinimide (6.2 g, 34.9 mmol) is added to a solution of1-methyl-1H-pyrrole-2-carbonitrile (3.7 g, 34.9 mmol) inN,N-dimethylformamide (150 mL). The solution is stirred for 15 hours atambient temperature. Another amount of N-bromosuccinimide (2.0 g, 11mmol) is added and the mixture is stirred for 3 hours. The product ispurified by chromatography over silica gel (ethyl acetate/heptanegradient) to obtain the expected product in the form of a solid.

¹H NMR (400 MHz, CDCl₃) δ ppm: 3.77 (s, 3H), 6.75 (d, J=1.7 Hz, 1H),6.80 (d, J=1.7 Hz, 1H)

Step C:4-[(tert-Butyldimethylsilyl)oxy]phenyl}amino)-1-methyl-1H-pyrrole-2-carbonitrile

Nitrogen is bubbled through a solution of4-bromo-1-methyl-1H-pyrrole-2-carbonitrile (2.82 g, 15.2 mmol) and4-[(tert-butyldimethylsilyl)oxy]aniline (4.08 g, 18.3 mmol) in toluene(55 mL) for 5 minutes. Sodium tert-butylate (2.92 g, 30.4 mmol),tris(dibenzylideneacetone)dipalladium(0) (556 mg, 0.6 mmol) and2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl (255 mg, 0.6mmol) are then added to the reaction mixture. The mixture is stirred for1 hour at 80° C. under nitrogen. The suspension is then cooled toambient temperature and filtered over Celite. The Celite cake is thenrinsed with ethyl acetate. The filtrate is washed with water and thenwith brine. The organic phase is dried over magnesium sulphate, filteredand concentrated in vacuo. The product is purified twice bychromatography over silica gel (AcOEt/heptane gradient), and then bytrituration in heptane to obtain the expected product in the form of asolid.

¹H NMR (400 MHz, CDCl₃) δ ppm: 0.16 (s, 6H), 0.97 (s, 9H), 3.73 (s, 3H),6.57 (d, J=1.9 Hz, 1H), 6.64-6.66 (m, 1H), 6.70 (s, 4H); NMR

¹³C NMR (100 MHz, CDCl₃) δ ppm: −4.48, 18.17, 25.72, 35.46, 103.01,113.56, 113.69, 115.92, 119.55, 120.67, 129.04, 139.94, 148.85

MS (ESI+): [M+H]⁺ 328.25

The amines NHR₃R₄ wherein R₃ and R₄, each independently of the other,represent an aryl or heteroaryl group are obtained in accordance withprocesses described in the literature (Surry D. S. et al., ChemicalScience, 2011, 2, 27-50, Charles M. D. et al., Organic Letters, 2005, 7,3965-3968). The reaction protecting the hydroxy function of the4-anilinophenol described in Preparation 1″ can be applied to varioussecondary amines NHR₃R₄ (as defined hereinbefore) having one or morehydroxy functions, when they are available commercially. Alternatively,the secondary amines having at least one hydroxy substituent may besynthesised directly in a protected form, i.e. starting from reagentswhose hydroxy function has been protected beforehand. Among theprotecting groups, tert-butyl(dimethyl)silyloxy and benzyloxy areespecially preferred.

Among the amines NHR₃R₄ having a hydroxy substituent that are used forsynthesising the compounds of the invention there may be mentioned:4-(4-toluidino)phenol, 4-(4-chloroanilino)phenol,4-(3-fluoro-4-methylanilino)phenol,4-[4-(trifluoromethoxy)anilino]phenol, 4-[4-hydroxyanilino]phenol,{4-[(1-methyl-1H-indol-6-yl)amino]phenyl}methanol,4-(2,3-dihydro-1H-indol-6-ylamino)phenol,4-[(1-methyl-2,3-dihydro-1H-indol-6-yl)amino]phenol,4-[(1-methyl-1H-indol-6-yl)amino]phenol,4-[(1-methyl-1H-indol-6-yl)amino]cyclohexanol,4-[(1-methyl-1,2,3,4-tetrahydro-6-quinolinyl)amino]phenol,4-[(4-methyl-3,4-dihydro-2H-1,4-benzoxazin-7-yl)amino]phenol,4-[4-(diethylamino)anilino]phenol,4-(2,3-dihydro-1H-inden-5-ylamino)phenol,4-[(1-methyl-1H-indazol-5-yl)amino]phenol,4-[(1′-methyl-1′,2′-dihydrospiro[cyclopropane-1,3′-indol]-5′-yl)amino]phenol,4-[(1,3,3-trimethyl-2,3-dihydro-1H-indol-5-yl) amino] phenol,4-[4-methoxy-3-(trifluoromethyl)anilino]phenol,4-[4-(methylsulphanyl)-3-(trifluoromethyl)anilino]phenol,2-fluoro-4-[(1-methyl-1H-indol-5-yl)amino]phenol,4-[(1-ethyl-1H-indol-5-yl)amino]phenol,4-[(1-ethyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,4-[(1-isopropyl-2,3-dihydro-1H-indol-5-yl)amino]phenol,4-(butylamino)phenol, 3-[(1-methyl-1H-indol-5-yl)amino]-1-propanol,4-[(1-methyl-1H-indol-5-yl)amino]-1-butanol,4-[(3-fluoro-4-methylphenyl)amino]phenol,4-[(3-chloro-4-methylphenyl)amino]phenol,4-[(4-fluorophenyl)amino]phenol,4-[(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenol,4-[(4-fluorophenyl)amino]phenol, 4-[(2-fluorophenyl)amino]phenol,4-[(3-fluorophenyl)amino]phenol, 4-[(2,4-difluorophenyl)amino]phenol,4-[(3,4-difluorophenyl)amino]phenol,3-[(4-hydroxyphenyl)amino]benzonitrile,4-[(3-methoxyphenyl)amino]phenol, 4-[(3,5-difluorophenyl)amino]phenol,4-[(3-methylphenyl)amino]phenol, 4-[(4-hydroxyphenyl)amino]benzonitrile,4-[(3-chlorophenyl)amino]phenol, 4-(pyrimidin-2-ylamino)phenol,4-[(cyclobutylmethyl)amino]phenol,2-[(4-hydroxyphenyl)amino]benzonitrile,4-{[(1-methyl-1H-pyrazol-4-yl)methyl]amino}phenol,4-[(cyclopropylmethyl)amino]phenol,4-{[(1-methyl-1H-pyrazol-3-yl)methyl]amino}phenol,4-(but-2-yn-1-ylamino)phenol, 4-(pyrazin-2-ylamino)phenol,4-(pyridin-2-ylamino)phenol, 4-(pyridazin-3-ylamino)phenol,4-(pyrimidin-5-ylamino)phenol, 4-(pyridin-3-ylamino)phenol,4-[(3,5-difluoro-4-methoxyphenyl)amino]phenol,4-(pyridin-4-ylamino)phenol, 4-[(3-fluoro-4-methoxyphenyl)amino]phenol,2-(phenylamino)pyrimidin-5-ol,5-[(4-hydroxyphenyl)amino]-2-methoxybenzonitrile and4-{[3-(trifluoromethyl)phenyl]amino}phenol.

The hydroxy function(s) of the secondary amines listed above is (are)protected beforehand by a suitable protecting group prior to anycoupling to an acid derivative of the compound of formula (VII) asdefined in the preceding general process.

EXAMPLE 14-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate Step A: Methyl3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)-carbonyl]-1,3-benzodioxol-5-yl}-5,6,7,8-tetrahydro-1-indolizine-carboxylate

To a solution of 2 g of the compound of Preparation 1 (5.83 mmol) in 20mL of dichloromethane there are added, at ambient temperature, 5.5 mL ofN,N,N-triethylamine (6.96 mmol), 2.12 g of the compound of Preparation1′ (6.96 mmol), and then 0.94 g of hydroxybenzotriazole (HOBT) and 1.34g of 1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (EDC) (6.96 mmol).The reaction mixture is then stirred at ambient temperature overnightand then it is poured into saturated aqueous ammonium chloride solutionand extracted with ethyl acetate. The organic phase is then dried overmagnesium sulphate and then filtered and evaporated to dryness. Thecrude product thereby obtained is then purified by chromatography oversilica gel (heptane/AcOEt gradient). The title product is obtained inthe form of an oil.

¹H NMR: δ (500 MHz; dmso-d6; 300° K): 7.2-6.9 (m, 4H, aromatic Hs);7.04-7.03-7.00 (m, 1H, aromatic H); 6.85 (m, 1H, aromatic H);6.35-6.26-6.06 (m, 1H, H tetrahydroindolizine); 6.15-6.12 (m, 2H, Hmethylenedioxy); 5.06-4.84 (m, 1H, H dihydroisoquinoline); 4.86-4.17 (m,2H, H dihydroisoquinoline); 3.65-3.6-3.55 (m, 3H, H methyl ester);3.43-4.26 (m, 2H, H tetrahydroindolizine); 3.58-3.5 (m, 4H, Hmorpholine); 2.37-3.05 (m, 4H, 2H dihydroisoquinoline, 2Htetrahydroindolizine); 1.68-2.56 (m, 4H, H morpholine); 1.4-2.0 (m, 4H,H tetrahydroindolizine)

IR: ν: >C═O 1695 cm⁻¹ ester; ν: >C═O 1625 cm⁻¹ amide; ν: >C—O—C<1214-1176-1115 cm⁻¹; >CH—Ar 772-744 cm⁻¹

Step B: Lithium3-[6-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-1,3-benzodioxol-5-yl]-5,6,7,8-tetrahydro-1-indolizine-carboxylate

To a solution of 4.6 g of the compound of Step A (8.26 mmol) in 24 mL ofdioxane there is added a solution of lithium hydroxide (675 mg, 16.1mmol). The batch is placed in a microwave oven at 140 W, 100° C. for aperiod of 2 hours 30 minutes. The reaction mixture is then filtered andevaporated. The solid thereby obtained is dried at 40° C. in an oven inthe presence of P₂O₅.

¹H NMR: δ (400 MHz; dmso-d6; 353° K): 6.7-7.15 (unresolved peak, 6H,aromatic Hs); 6.21 (s, 1H, aromatic H); 6.03 (s, 2H, H methylenedioxy);4.0-5.0 (unresolved peak, 3H dihydroisoquinoline); 3.4-3.6 (unresolvedpeak, 3H tetrahydroindolizine, 3H morpholine); 2.5-3.1 (unresolved peak,4H, 2H tetrahydroindolizine, 2H morpholine); 1.5-2.4 (unresolved peak,10H morpholine)

IR: ν: >C═O broad 1567 cm⁻¹ acetate; ν: 1236 cm⁻¹

Step C:N-(4-{[tert-Butyl(dimethyl)silyl]oxy}phenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide

To a solution of 2.6 g of the compound of Step B (4.73 mmol) in 47 mL ofdichloromethane there are added, dropwise, 1.2 mL of oxalyl chloride(14.2 mmol) at 0° C. The reaction mixture is stirred at ambienttemperature for 11 hours and then co-evaporated several times withdichloromethane. The product thereby obtained is suspended in 37 mL ofdichloromethane, and is then added to a solution of 2.1 g of thecompound obtained in Preparation 1″ (7.1 mmol) in 10 mL ofdichloromethane in the presence of 0.6 mL of pyridine (7.1 mmol). Thebatch is stirred at ambient temperature overnight.

The reaction mixture is concentrated and purified by chromatography oversilica gel (dichloromethane/methanol gradient). The title product isobtained in the form of a foam.

¹H NMR: δ (500MHz; dmso-d6; 300° K): 6.9-7.3 (9H, aromatic Hs); 6.88(2H, aromatic Hs); 6.72-6.87 (2H, aromatic Hs); 6.64 (2H, aromatic Hs);6.13 (2H methylenedioxy); 5.05-4.74 (1H dihydroisoquinoline); 4.25-4.13(2H dihydroisoquinoline); 3.44-3.7 (4H morpholine); 3.62-3.52 (2Htetrahydroindolizine); 3.0-2.6 (4H, 2H tetrahydroindolizine, 2Hdihydroisoquinoline); 2.54-1.94 (6H morpholine); 1.91-1.53 (4Htetrahydroindolizine); 0.92 (9H tert-butyl); 0.17 (6H dimethyl)

IR: ν: >C═O: 1632 cm⁻¹; ν: >C—O—C<: 1237 cm⁻¹; ν: —Si—O—C—: 1035 cm⁻¹;—Si—C—: 910 cm⁻¹; >CH—Ar: 806 cm⁻¹

Step D:N-(4-Hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide hydrochloride

To a solution of 1.9 g of the compound obtained in Step C (2.3 mmol) in4 mL of methanol there is added 0.646 g (11.5 mmol) of potassiumhydroxide dissolved in 8 mL of methanol. The batch is stirred at ambienttemperature for 30 minutes. The reaction mixture is then diluted withdichloromethane and washed successively with 1M HCl solution, saturatedaqueous NaHCO₃ solution and then brine until a neutral pH is reached.The organic phase is then dried over magnesium sulphate, filtered andevaporated. The crude product thereby obtained is purified bychromatography over silica gel (dichloromethane/methanol gradient). Thesolid is then dissolved in dichloromethane, and 2 mL of 1M ethereal HClare added. The batch is stirred for 1 hour and then evaporated todryness. The hydrochloride thereby obtained is dissolved in a mixture ofwater/acetonitrile until dissolution is complete, and is thenlyophilised.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.11:68.95; % H=5.8:5.46; % N=7.5:7.51; % Cl-=4.74:4.48

Optical rotation: (α)_(D) ²⁰+50.8° (c=9 mg/mL, MeOH)

Step E:4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-ylicarbonyl}(phenyl)amino]phenyldibenzyl phosphate

To a suspension of 82 mg of sodium hydride (2.06 mmol) in 10 mL ofanhydrous THF there are added, in portions and at 0° C., 700 mg of thecompound of Step D. After stirring for 30 minutes at 0° C. and for 30minutes at ambient temperature, tetrabenzyl pyrophosphate is added at 0°C. and the reaction mixture is stirred overnight at ambient temperature.After evaporating off the solvent, the crude reaction product is dilutedwith dichloromethane (30 mL), washed with saturated aqueous NaHCO₃solution and then with brine. The organic phase is then dried overMgSO₄, filtered, concentrated to dryness and purified by chromatographyover silica gel (gradient CH₂Cl₂/MeOH). The title product is thenobtained in the form of a solid.

¹H NMR: δ (500 MHz; DMSO-d6; 300K): 7.34 (m, 10H, phenyl); 7.30-6.71 (m,15H, aryl); 6.06 (s, 1H, methylenedioxy); 5.30-4.97 (m, 1H, pyrrole);5.11 (m, 4H, benzyl): 5.03-3.64 (m, 1H, tertiary C THIQ); 4.91-4.09 (m,2H, secondary C THIQ); 3.99-3.48 (m, 2H, secondary C THIQ); 3.54-3.44(m, 4H, morpholine); 2.89-2.65 (m, 3H, secondary C THIQ); 2.51-1.87 (m,4H, secondary C THID); 2.36-1.85 (m, 2H, secondary C THIQ); 1.91-1.45(m, 4H, secondary C THID)

Step F:4-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate

50 mg of Pd(OH)₂ are added to a solution of the product obtained in StepE (505 mg; 0.52 mmol) in methanol (10 mL), and then the reaction mixtureis placed under a hydrogen atmosphere (1 bar) for 5 hours. Afterfiltering off the catalyst and concentrating to dryness, the crudereaction product is dissolved in methanol (5 mL) and treated with 0.95mL of 1M sodium hydroxide solution. The solvents are then evaporated offand the crude reaction product is purified by chromatography over anOASIS® phase (acetonitrile/H₂O gradient) to obtain a white solid.

Elemental Microanalysis:

% C % H % N % Na Calculated 61.87 4.95 6.71 5.51 Found 61.45 4.46 6.615.38

IR: ν: —C═O: 1628 cm⁻¹; ν: C—O—C: 1234 cm⁻¹; ν: P═O: 115 cm⁻¹; ν: P—O:985 cm⁻¹; ν: CH—Ar: 876 cm⁻¹

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₃H₄₁N₄Na₂O₉P

[M+H]+ calculated: 835.2479

[M+H]+ measured: 835.2467

EXAMPLE 24-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-phenyl-5,6,7,8-tetrahydroindolizine-1-carboxamide

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing the product of Preparation1′ in Step A by that of Preparation 2′, it being understood that theproduct thereby obtained is not subjected to a step of conversion intosalt form in the presence of HCl in ether.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=74.86:74.88; % H=5.64:5.31; % N=6.72:6.78

Step B:4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps E and F of Example 1.

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₉H₃₆N₃O₈P

[M+H]⁺ calculated: 706.2313

[M+H]+ measured: 706.2324

EXAMPLE 34-[(1-Methyl-1H-indol-5-yl){[3-(2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}amino]phenyldisodium phosphate Step A:3-{5-Chloro-2-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]phenyl}-N-(4-hydroxyphenyl)-N-(1-methyl-1H-indol-5-yl)-5,6,7,8-tetrahydro-1-indolizinecarboxamide hydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation2 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 2″.

Elemental Microanalysis:

% C % H % N % Cl⁻ Calculated 68.04 5.72 8.82 4.91 Found 67.84 5.46 8.645.21

Optical rotation: (α)_(D) ²⁰+55.9° (c=7 mg/mL, MeOH)

Step B:4-[(1-Methyl-1H-indol-5-yl){[3-(2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps E and F of Example 1.

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₅H₄₄N₅Na₂O₇P

[M−2Na+3H]⁺ calculated: 800.3208

[M−2Na+3H]+ measured: 800.3211

EXAMPLE 44-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-indolizine-1-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompounds of Preparations 1 and 1′ used in Step A by the compounds ofPreparations 3 and 2′ and, on the other hand, the compound ofPreparation 1″ used in Step C by that of Preparation 3″.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.14:70.09; % H=4.81:4.55; % N=9.83:10.09; % Cl-=4.98:3.26

Step B:4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldiethyl phosphate

To a suspension of the compound obtained in Step A (1.5 mmol) in 10 mLof anhydrous CH₂Cl₂ there are added triethylamine (0.42 mL; 3 mmol), andthen diethyl cyanophosphonate (0.24 mL; 1.65 mmol) dropwise at ambienttemperature. After stirring overnight at ambient temperature, thereaction mixture is diluted with CH₂Cl₂, washed with saturated aqueousNaHCO₃ solution and then with brine. The organic phase is then driedover MgSO₄, filtered, concentrated to dryness and purified bychromatography over silica gel (CH₂Cl₂/MeOH gradient). The title productis then obtained in the form of a solid.

Step C:4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldisodium phosphate

0.4 mL of trimethylsilyl bromide (3 mmol) is added dropwise at ambienttemperature to a solution of the product obtained in Step B (0.78 mmol)in CH₂Cl₂ (12 mL). The reaction mixture is stirred for 5 hours, and thena solution of Na₂CO₃ (580 mg) in water (4 mL) is slowly added at 0° C.After stirring for 30 minutes, the reaction mixture is concentrated todryness, diluted with anhydrous methanol (25 mL) and microfiltered. Thefiltrate is dried and purified by chromatography over an OASIS® phase(acetonitrile/H₂O gradient).

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₅H₄₄N₅Na₂O₇P

[M−2Na+3H]⁺ calculated: 800.3211

[M−2Na+3H]+ measured: 800.3201

EXAMPLE 54-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-N-(pyridin-4-yl)indolizine-1-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompounds of Preparations 1 and 1′ used in Step A by the compounds ofPreparations 3 and 2′ and, on the other hand, the compound ofPreparation 1″ used in Step C by that of Preparation 4″.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=69.24:69.12; % H=4.74:4.23; % N=8.5:8.45; % Cl-=5.38:5.2

Step B:4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyldiethyl phosphate

To a suspension of 950 mg of the compound obtained in Step A (1.5 mmol)in 10 mL of anhydrous CH₂Cl₂ there are added triethylamine (0.42 mL; 3mmol), and then diethyl cyanophosphonate (0.24 mL; 1.65 mmol) dropwiseat ambient temperature. After stirring overnight at ambient temperature,the reaction mixture is diluted with CH₂Cl₂, washed with saturatedaqueous NaHCO₃ solution and then with brine. The organic phase is thendried over MgSO₄, filtered, concentrated to dryness and purified bychromatography over silica gel (CH₂Cl₂/MeOH gradient). The title productis then obtained in the form of a solid.

¹H NMR: δ (500 MHz; DMSO-d6; 300K): 8.5-8. 0 (m, 5H); 7.2-7.1 (m, 1H);6.85-6.65 (m, 1H); 7.3-6.8 (m, 10H); 6.25-6.10 (bs, 1H); 6.2 (bs, 2H);5.1-3.7 (6d, 2H); 4.7-3.8 (m,1H); 4.15 (m, 4H); 3.0-1.7 (m,2H); 1.25 (m,6H); 0.85-0.24 (m,3H)

Step C:4-[{[3-(6-{[(3R)-3-Methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate

0.4 mL of trimethylsilyl bromide (3 mmol) is added dropwise at ambienttemperature to a solution of the product obtained in Step B (591 mg;0.78 mmol) in CH₂Cl₂ (12 mL). The reaction mixture is stirred for 5hours, and then a solution of Na₂CO₃ (580 mg) in water (4 mL) is slowlyadded at 0° C. After stirring for 30 minutes, the reaction mixture isconcentrated to dryness, diluted with anhydrous methanol (25 mL) andmicrofiltered. The filtrate is dried and purified by chromatography overan OASIS® phase (acetonitrile/H₂O gradient).

¹H NMR: δ (500 MHz; D₂O; 300K): 8.23-7.98 (m, 2H, pyridyl); 7.01-6.97(m, 2H, pyridyl); 7.88-7.80 (m, 1H, indolizine); 7.18-6.57 (m, 13H,aromatic Hs THIQ+aryl+indolizine+phenol); 6.17-6.15 (m, 1H, indolizine):5.96 (m, 2H, methylenedioxy); 4.61-3.76 (m, 1H, ternary C THIQ); 4.16(m, 2H, secondary C THIQ); 2.86-2.31 (m, 2H, secondary C THIQ);0.94-0.76 (m, 3H, primary C THIQ)

IR: ν: —C═O: 1620 cm⁻¹; ν: C—O—C: 1218 cm⁻¹; ν: P═O: 1107 cm⁻¹ ν: P—O:981 cm⁻¹ ν: CH—Ar: 881-741 cm⁻¹

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₈H₂₉N₄Na₂O₈P

[M−2Na+3H]⁺ calculated: 703.1952

[M−2Na+3H]+ measured: 703.1951

EXAMPLE 64-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyl dibenzyl phosphate

The procedure is in accordance with the protocol described in Steps A-Eof Example 1.

¹H NMR: δ (500 MHz; DMSO-d6; 300K): 7.34 (m, 10H, phenyl); 7.30-6.71 (m,15H, aryl); 6.06 (s, 1H, methylenedioxy); 5.30-4.97 (m, 1H, pyrrole);5.11 (m, 4H, benzyl): 5.03-3.64 (m, 1H, tertiary C THIQ); 4.91-4.09 (m,2H, secondary C THIQ); 3.99-3.48 (m, 2H, secondary C THIQ); 3.54-3.44(m, 4H, morpholine); 2.89-2.65 (m, 3H, secondary C THIQ); 2.51-1.87 (m,4H, secondary C THID); 2.36-1.85 (m, 2H, secondary C THIQ); 1.91-1.45(m, 4H, secondary C THID)

EXAMPLE 7 Diethyl4-[{[3-(6-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)indolizin-1-yl]carbonyl}(pyridin-4-yl)amino]phenylphosphate

The procedure is in accordance with the protocol described in Steps Aand B of Example 5.

¹H NMR: δ (500 MHz; DMSO-d6; 300K): 8.5-8. 0 (m, 5H); 7.2-7.1 (m, 1H);6.85-6.65 (m, 1H); 7.3-6.8 (m, 10H); 6.25-6.10 (bs, 1H); 6.2 (bs, 2H);5.1-3.7 (6d, 2H); 4.7-3.8 (m, 1H); 4.15 (m, 4H); 3.0-1.7 (m,2H); 1.25(m, 6H); 0.85-0.24 (m, 3H)

EXAMPLE 84-[{[3-(6-{[(3S)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldihydrogen phosphate hydrochloride

100 mg of Pd(OH)₂ are added to a solution of the product obtained inStep E of Example 1 (500 mg; 0.51 mmol) in methanol (10 mL), and thenthe reaction mixture is placed under a hydrogen atmosphere (1 bar) for 5hours. After filtering off the catalyst and concentrating to dryness,the crude reaction product is immediately purified by chromatographyover a C18 phase (acetonitrile/H₂O+0.2% HCl gradient) to obtain a solid.

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₃H₄₃N₄O₉P

[M+H]⁺ calculated: 791.2846

[M+H]⁺ measured: 791.2852

EXAMPLE 94-[{[5-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate Step A:5-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(pyridin-4-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompounds of Preparations 1 and 1′ used in Step A by the compounds ofPreparations 4 and 2′ and, on the other hand, the compound ofPreparation 1″ used in Step C by that of Preparation 4″.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=66.99:66.88; % H=5.14:5.28; % N=8.93:8.87; % Cl-=5.65:4.98

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₅H₃₂ClN₄O₃

[M+H]⁺ calculated: 591.2157

[M+H]⁺ measured: 591.2178

Step B:4-[{[5-(5-Chloro-2-{[(3R)-3-methyl-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 4.

EXAMPLE 104-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation5 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 3″.

Elemental Microanalysis: (%, Measured(Theoretical))

% C=66.41(66.62); % H=5.08(5.59); % N=10.85(10.84); % Cl-=4.68(4.57)

Step B:4-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 4.

EXAMPLE 114-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrazol-4-yl)-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation5 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 5″.

Elemental Microanalysis: (%, Measured(Theoretical))

% C=64.25(64.59); % H=5.4(5.7); % N=11.41(11.59); % Cl-=4.93(4.89)

Step B:4-[{[1,2-Dimethyl-5-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps E and F of Example 1.

IR (cm⁻¹): ν: C═O: 1628; ν: (phosphate; ether): 1238, 1143,1113, 985;γ: >CH Ar: 740

Elemental Microanalysis:

% C % H % N Calculated 57.64 4.84 10.34 Found 56.62 4.54 10.14

High-Resolution Mass Spectrometry (ESI+−/FIA/HR):

Empirical formula: C₃₉H₃₉ClN₆Na₂O₉P

[M−Na+H]⁺ calculated: 791.2565

[M−Na+H]⁺ measured: 791.2564

EXAMPLE 144-[{[1,2-Dimethyl-5-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)-5-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompounds of Preparations 1 and 1′ used in Step A by the compounds ofPreparations 5 and 3′ and, on the other hand, the compound ofPreparation 1″ used in Step C by that of Preparation 3″.

Elemental Microanalysis: (%, Measured(Theoretical))

% C=67.63(68.06); % H=5.27(5.95); % N=10.08(10.13); % Cl-=4.53(4.27)

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₅H₃₂ClN₄O₃

[M+H]⁺ calculated: 793.3708

[M+H]⁺ measured: 793.3704

Step B:4-[{[1,2-Dimethyl-5-(6-{[(3R)-3-[3-(morpholin-4-yl)propyl]-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrrolo[2,3-b]pyridin-5-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps E and F of Example 1.

Unless otherwise mentioned, the compounds of the following Examples aresynthesised in accordance with the process of Example 1 using: (i) theappropriate acid obtained according to one of Preparations 1 to 9 and(ii) the appropriate tetrahydroisoquinoline compound obtained accordingto one of Preparations 1′ to 4′ and, in Step C: (iii) the suitableNHR₃R₄ amine (a non-exhaustive list is proposed in Preparations 1″ to8″).

EXAMPLE 134-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate Step A:5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(pyridin-4-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation4 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 4″. The product obtained is finally subjected toa step of conversion into salt form in the presence of 1M HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₉H₃₈ClN₅O₄

[M+H]⁺ calculated: 676.2685

[M+H]⁺ measured: 676.2684

Step B:4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (500 MHz, D₂O) δ ppm: −0.05

IR (cm⁻¹): ν: C═O: 1631; ν: (phosphate; ether): 1243, 1136, 1112, 982;γ: >CH Ar: 883, 745

Elemental Microanalysis:

% C % H % N Calculated 58.54 4.66 8.75 Found 58.23 4.51 8.76

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₉H₃₇ClN₅Na₂O₇P

[M−Na+2H]⁺ calculated: 778.2168

[M−Na+2H]⁺ measured: 778.2169

EXAMPLE 144-[{[5-(5-Fluoro-4-methoxy-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate EXAMPLE 154-[{[5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate Step A:5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation7 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 5″. The product obtained is finally subjected toa step of conversion into salt form in the presence of HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

Elemental Microanalysis: (%, Measured(Theoretical))

% C=65.69(65.28); % H=5.38(5.77); % N=11.18(12.02); % Cl-=5.61(5.07)

Step B:4-[{[5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (400/500 MHz, CD₃OD) δ ppm: −0.5

IR (cm⁻¹): ν: C═O: 1628; ν: (phosphate; ether): 1238, 1114, 983

Elemental Microanalysis:

% C % H % N Calculated 58.02 4.87 10.68 Found 59.03 4.98 10.14

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₈H₃₈FN₆Na₂O₇P

[M−2Na+3H]⁺ calculated: 743.2752

[M−2Na+3H]⁺ measured: 743.2760

EXAMPLE 164-[{[1,2-Dimethyl-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrazol-4-yl)-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation8 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 5″. The product obtained is finally subjected toa step of conversion into salt form in the presence of 1M HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=64.99:64.67; % H=5.86:5.67; % N=11.37:11.27; % Cl-=4.8:4.71

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₀H₄₃N₆O₆

[M+H]⁺ calculated: 703.3236

[M+H]⁺ measured: 703.3239

Step B:4-[{[1,2-Dimethyl-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenylN,N,N′,N′-tetramethylphosphorodiamidate

To a solution of 125 mg of the compound of Step A (0.18 mmol) indichloromethane (6 mL) there are added 55 μL ofdiazabicyclo[5.4.0]undec-7-ene (DBU; 0.36 mmol), and then 33 μL ofbisdimethylaminophosphoryl chloride (0.19 mmol) and 2 mg ofdimethylamino-4-pyridine (0.02 mmol). The reaction mixture is stirredfor 15 hours, diluted with dichloromethane and then with saturatedaqueous sodium carbonate solution. The aqueous phase is extracted withdichloromethane; the organic phases are then combined, washed with waterand with brine and dried over magnesium sulphate. After evaporating offthe solvents, the crude reaction product is used directly in the nextStep.

Step C:4-[{[1,2-Dimethyl-5-(7-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-2,3-dihydro-1,4-benzodioxin-6-yl)-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate

4 mL of trifluoroacetic acid are added dropwise to a solution of 125 mgof the compound of Step B (0.15 mmol) in a 1:1 mixture of acetonitrileand water (5 mL). After stirring for 20 hours at ambient temperature,the reaction mixture is evaporated to dryness, keeping the temperatureof the water bath below 40° C., and then the residue is treated with asolution of sodium carbonate (95 mg; 0.9 mmol) in water (4 mL). Afterstirring for 2 hours at ambient temperature, the reaction mixture isevaporated to dryness and then 6 mL of anhydrous ethanol are added. Thesolid is filtered off, and the filtrate is concentrated to dryness, andthen purified over an OASIS® phase (acetonitrile/water gradient).

³¹P NMR (500 MHz, D₂O) δ ppm: 0.9

IR (cm⁻¹): ν: C═O: 1623; ν: (phosphate; ether): 1235, 1162, 1115, 1065,985; γ: >CH Ar: 745

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₀H₄₁N₆Na₂O₉P

[M−2Na+3H]⁺ calculated: 783.2902

[M−2Na+3H]⁺ measured: 783.2907

EXAMPLE 175-[{[5-(5-Fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]pyrimidin-2-yldisodium phosphate EXAMPLE 185-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]pyrimidin-2-yldisodium phosphate EXAMPLE 194-({[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideuteriomethyl)-1H-pyrazol-4-yl]amino)phenyldisodium phosphate Step A:5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-[1-(trideuteriomethyl)-1H-pyrazol-4-yl]-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation4 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 6″. The product obtained is finally subjected toa step of conversion into salt form in the presence of 1M HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=63.51:63.41; % H=5.63:5.42; % N=11.69:11.61; % Cl⁻=4.93:4.85

High-Resolution Mass Spectrometry (ESI+−/FIA/HR, ESI−/FIA):

Empirical formula: C₃₈H₃₆ClD₃N₆O₄

[M+H]⁺ calculated: 682.2982

[M+H]⁺ measured: 682.2986

Step B:4-({[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideuteriomethyl)-1H-pyrazol-4-yl]amino)phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (500 MHz, D₂O) δ ppm: 4.8

IR (cm⁻¹): ν: C═O: 1626; ν: (phosphate; ether): 1243, 1141, 1115, 982;γ: >CH Ar: 880, 831

High-Resolution Mass Spectrometry (ESI/FIA/HR and MS/MS):

Empirical formula: C₃₈H₃₅ClD₃N₆Na₂O₇P

[M+H]⁺ calculated: 806.2285

[M+H]⁺ measured: 806.2280

EXAMPLE 204-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)amino]phenyldisodium phosphate Step A:5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)-N-(4-hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation4 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 7″. The product obtained is finally subjected toa step of conversion into salt form in the presence of 1M HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

¹H NMR (500 MHz, dmso-d6) δ ppm: 11.2 (bs, 1H), 9.39 (bs,1H), 7.83 (d,1H), 7.54 (d, 1H), 7.33 (s, 1H), 7.14 (m, 2H), 7 (m, 2H), 6.8 (d, 2H),6.62 (d, 2H), 6.57 (bs, 1H), 5.26 (s, 1H), 5.26 (m, 1H), 4.64/4.03 (AB,2H), 4.01/3.92 (2m, 4H), 3.75/3.43/3.15/3.02 (4m, 4H), 3.59 (s, 3H),3.3/3.15 (2m, 2H), 2.97 (s, 3H), 2.69/2.52 (dd+d, 2H), 2.06 (s, 3H),1.91 (s, 3H)

Elemental Microanalysis: (%, Theoretical:Measured)

% C=65.34:65.50; % H=5.62:5.15; % N=11.15:10.84% Cl-=4.70:4.44

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₁H₄₁ClN₆O₄

[M+H]⁺ calculated: 717.2952

[M+H]⁺ measured: 717.2951

Step B:4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (500 MHz, dmso-d6) δ ppm: 3.7

IR (cm⁻¹): ν: —CN: 2210 cm⁻¹; ν: C═O: 1623; ν: (phosphate; ether): 1227,1133, 1110, 982; γ: >CH Ar: 884-741

Elemental Microanalysis:

% C % H % N Calculated 58.54 4.79 9.99 Found 58.75 4.71 10.18

High-Resolution Mass Spectrometry (ESI+−/FIA/HR):

Empirical formula: C₄₁H₄₀ClN₆Na₂O₇P

[M−2Na+H]⁺ calculated: 797.2614

[M−2Na+H]⁺ measured: 797.2618

EXAMPLE 214-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-methyl-1H-pyrrol-3-yl)amino]phenyldisodium phosphate Step A:5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(5-cyano-1-methyl-1H-pyrrol-3-yl)-N-(4-hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation4 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 8″. The product obtained is finally subjected toa step of conversion into salt form in the presence of 1M HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=64.95:65.09; % H=5.45:5.20; % N=11.36:11.26; % Cl-=4.79:4.62

High-Resolution Mass Spectrometry (ESI/+):

Empirical formula: C₄₀H₃₉ClN₆O₄

[M+H]⁺ calculated: 703.2794

[M+H]⁺ measured: 703.2789

Step B4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-methyl-1H-pyrrol-3-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (500 MHz, dmso-d6) δ ppm: 4.5

IR (cm⁻¹): ν: —CN: 2215 cm⁻¹; ν: C═O 1626; ν: (Phosphate; ether): 1227,1141, 1112, 982; γ >CH Ar: 826-742

High-Resolution Mass Spectrometry (ESI+−/FIA/HR):

Empirical formula: C₄₀H₃₈ClN₆Na₂O₇P

[M−2Na+3H]⁺ calculated: 783.2457

[M−2Na+3H]⁺ measured: 783.2462

EXAMPLE 224-[{[3-(6-{[(3R)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate Step A:N-(4-Hydroxyphenyl)-3-{6-[((3R)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide hydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing the compound ofPreparation 1′ used in Step A by the compound of Preparation 4′. Thesolid is then dissolved in dichloromethane, and 2 mL of 1M HCl in etherare added. The batch is stirred for 1 hour and then evaporated todryness. The hydrochloride thereby obtained is dissolved in a mixture ofwater/acetonitrile until dissolution is complete, and then lyophilised.

Elemental Microanalysis:

% C % H % N % Cl⁻ Calculated 69.11 5.80 7.50 4.74 Found 68.89 5.23 7.414.62

Optical rotation: (α)_(D) ²⁰=−45.1° (c=9 mg/mL, MeOH)

Step B:4-[{[3-(6-{[(3R)-3-(Morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (400/500 MHz, dmso-d6) δ ppm: 2.6

Elemental Microanalysis:

% C % H % N Calculated 61.87 4.95 6.71 Found 61.33 4.93 7.14

High-Resolution Mass Spectrometry (ESI+−/FIA/HR):

Empirical formula: C₄₃H₄₁N₄Na₂O₉P

[M−2Na+H[⁺ calculated: 791.2840

[M−2Na+H[⁺ measured: 791.2845

EXAMPLE 234-[(1-Methyl-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-5-yl){[3-(2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-4-[2-oxo-2-(piperidin-1-yl)ethoxy]phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}amino]phenyldisodium phosphate Step A: Methyl3-[4-benzyloxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxylate

To a solution of 14.19 g (35.0 mmol) of the compound obtained inPreparation 9 in 200 mL of dimethylformamide there are successivelyadded 4-[[(3S)-1,2,3,4-tetrahydroisoquinolin-3-yl]methyl]morpholine(Preparation 1′; 8.13 g; 35.0 mmol), hydroxybenzotriazole (6.15 g; 45.5mmol), 1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide hydrochloride(6.70 g; 45.5 mmol) and triethylamine (21.95 mL; 0.16 mol). The batch isthen stirred overnight at ambient temperature. The reaction mixture isthen poured into 400 mL of ethyl acetate and then successively washedwith saturated aqueous NaHCO₃ solution, water and brine. The combinedaqueous phases are extracted with ethyl acetate. The resulting organicphases are dried over sodium sulphate, filtered and concentrated underreduced pressure. The product obtained is purified by chromatographyover silica gel to provide the title compound.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 7.5-7.3 (m, 5H), 7.38 (d, 1H),7.2-6.9 (m, 4H), 7.15 (dd, 1H), 6.9 (d, 1H), 6.35/6.25/6.08 (3*s, 1H),5.21/5.12 (3*s, 2H), 5.09/4.85/3.7 (3*m, 1H), 4.9-3.8 (8*d, 2H), 4.2-3.4(m, 2H), 3.65/3.6/3.55 (3*s, 3H), 3.6-3.4 (m, 4H), 3-2.4 (m, 2H),2.9-1.8 (6*dd, 2H), 2.5-1.95 (4*m, 4H), 2.35-1.7 (6*m, 2H), 2-1.45 (6*m,4H)

Step B:3-[4-Benzyloxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxylicacid

40.1 mL of 1M aqueous LiOH solution are added to a solution of 12.7 g(20 mmol) of the compound obtained in the Step above in 40 mL ofdioxane. The batch is heated at 100° C. overnight. The reaction mixtureis poured into water and then extracted with ethyl ether. The etherealphase is extracted once more with water. The combined aqueous phases areacidified to pH 4 by adding powdered citric acid, and then extractedwith dichloromethane. The dichloromethane phase is washed with brine,dried over sodium sulphate, filtered and concentrated to dryness. Thetitle product is obtained in the form of a meringue.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 11.35 (bs, 1H), 7.5-7.3 (m, 5H),7.38 (m, 1H), 7.2-6.9 (m, 4H), 7.15 (m, 1H), 6.9 (m, 1H), 6.31/6.25/6.1(3*s, 1H), 5.22/5.2/5.15 (3*s, 2H), 5.1/4.82/3.7 (3*m, 1H), 4.85-3.8(8*d, 2H), 4.2-3.4 (m, 2H), 3.6-3.45 (m, 4H), 3-2.3 (m, 2H), 2.9-1.8 (m,2H), 2.5-1.9 (6*m, 4H), 2.35-1.8 (6*m, 2H), 1.9-1.3 (m, 4H)

Step C:3-[4-Benzyloxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]phenyl]-N-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-N-(1-methylpyrrolo[2,3-b]pyridin-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

The acid obtained in Step B (9 g, 11.8 mmol) is dissolved in 90 mL of1,2-dichloroethane. 1.9 mL of 1-chloro-N,N,2-trimethylpropenylamine (14mmol) are added thereto. After stirring for 3 hours at ambienttemperature, 90 mL of toluene and 4.62 g ofN-[4-[tert-butyl(dimethyl)silyl]oxyphenyl]-1-methyl-1H-pyrrolo[2,3-b]pyridin-5-amine(Preparation 3″, 13 mmol) are added. The reaction mixture is heated at110° C. for 20 hours. After returning to ambient temperature, thereaction mixture is washed with brine, dried over Na₂SO₄, filtered andconcentrated to dryness. The residue obtained is purified bychromatography over silica gel (dichloromethane/ethanol gradient) toyield the expected product.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 7.95/7.8/7.75 (3*d, 1H),7.68/7.65/7.4 (3*d, 1H), 7.4/7.3 (2*d, 1H), 7.25-6.8 (m, 9H), 7.05/6.9(2*m, 1H), 7-6.6 (3*bd, 2H), 6.9 (m, 1H), 6.75-6.45 (3*bd, 2H), 6.7 (m,1H), 6.3 (2*d, 1H), 5.15-4.95 (m, 2H), 5.15/5.1/4.8 (3*s, 1H),4.95/4.6/3.5 (3*m, 1H), 4.9-3.7 (8*d, 2H), 3.8-3.3 (3*m, 2H),3.75/3.7/3.5 (3*s, 3H), 3.45/3.3 (2*m, 4H), 3-2.5 (3*m, 2H), 3-2.3 (m,2H), 2.4-1.75 (5*m, 4H), 2.25-1.7 (6*m, 2H), 1.75-1.3 (m, 4H), 0.7 (bs,9H), 0.1 (m, 6H)

Step D:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-3-[4-hydroxy-2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]phenyl]-N-(1-methylpyrrolo[2,3-b]pyridin-5-yl)-5,6,7,8-tetrahydroindolizine-1-carboxamide

0.9 g of Pd/C 10% is added to a solution, in 100 mL of ethanol, of 8.88g (8.4 mmol) of the compound obtained in Step C, whilst bubbling throughargon. The reaction mixture is placed under 1.2 bars of hydrogen atambient temperature for 15 hours. The catalyst is filtered off and thesolvent is evaporated off under reduced pressure to provide the titlecompound.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 8.06/7.92/7.87 (3*d, 1H),7.75/7.5/7.39 (3*d, 1H), 7.5 (m, 1H), 7.28-6.9 (m, 5H), 6.87/6.7 (2*m,2H), 6.76 (m, 1H), 6.75/6.67/6.62 (3*m, 2H), 6.67/6.46 (m, 1H), 6.4/6.36(2*m, 1H), 5.19/5.13/4.9 (3*bs, 1H), 5.06/4.7/3.6 (3*m, 1H),4.97/4.2/4.15/4.07 (4*m, 2H), 4.87/4.81 (bs, 1H), 3.86/3.56/3.39 (3*m,2H), 3.78/3.57 (2*m, 3H), 3.59/3.44 (2*m, 4H), 2.96-2.61 (2*m, 2H),2.88/2.6 (2*m, 2H), 2.59-1.81 (m, 6H), 1.87-1.42 (m, 4H), 0.89 (s, 9H),0.12 (m, 6H)

Step E:N-[4-[tert-Butyl(dimethyl)silyl]oxyphenyl]-N-(1-methylpyrrolo[2,3-b]pyridin-5-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxamide

The compound of Step D (3.0 g, 2.9 mmol) is dissolved in 100 mL oftoluene. 1.53 g (5.8 mmol) of triphenylphosphine and 0.62 g (4.3 mmol)of 2-hydroxy-1-(1-piperidyl)ethanone are added thereto. The mixture isheated to 50° C., and then 1.01 g (4.3 mmol) of di-tert-butylazodicarboxylate are added. The reaction mixture is stirred at 50° C.for 1 hour and is then allowed to return to ambient temperature beforeadding 1 mL of trifluoroacetic acid. After stirring overnight at ambienttemperature, the mixture is successively washed with water, saturatedNaHCO₃ and brine solution. The combined aqueous phases are extractedwith ethyl acetate. The resulting organic phases are dried over sodiumsulphate, filtered and concentrated under reduced pressure. The crudeproduct obtained is purified by chromatography over silica gel(dichloromethane/ethanol 98/2) to yield the expected compound.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 8.06/7.92/7.87 (3*d, 1H),7.75/7.51/7.4 (3*d, 1H), 7.49 (2*d, 1H), 7.29-6.89 (m, 5H), 6.93 (m,1H), 6.88/6.7 (m, 2H), 6.75/6.67 (m, 1H), 6.75/6.68/6.59 (3*m, 2H),6.4/6.36 (2*m, 1H), 5.2/5.16/4.92 (3*m, 1H), 5.06/4.69/3.58 (3*m, 1H),4.97/4.25/4.16/4.03 (4*d, 2H), 4.89/4.81 (2*m, 2H), 3.79/3.59 (2*m, 3H),3.59/3.43/3.4 (3*m, 6H), 3.58/3.43 (2*m, 4H), 3.03-2.61 (m, 2H),2.97-2.65 (m, 2H), 2.57-1.74 (m, 6H), 1.89-1.3 (m, 10H), 0.89 (2bs, 9H),0.11 (m, 6H)

Step F:N-(4-Hydroxyphenyl)-N-(1-methylpyrrolo[2,3-b]pyridin-5-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxamide

A 1M solution of tetrabutylammonium fluoride (3.14 mL, 3 mmol) intetrahydrofuran is added at ambient temperature to a solution, in 30 mLof tetrahydrofuran, of the compound obtained in Step E (2.92 g, 2.9mmol). After stirring for 5 minutes, the reaction mixture is poured intoa 50/50 mixture of ethyl acetate and saturated aqueous NaHCO₃ solution.The separated organic phase is washed with water and then with brine.The combined aqueous phases are extracted with ethyl acetate. Theresulting organic phases are dried over sodium sulphate, filtered andconcentrated under reduced pressure. The crude product obtained ispurified by chromatography over silica gel(dichloromethane/ethanol/ammonia gradient) to yield the title compound.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 9.4 (m, OH), 8.1-7.8 (3*d, 1H),7.7-7.3 (2*m, 1H), 7.5/7.4 (2*m, 1H), 7.3-6.9 (m, 4H), 7.2 (m, 1H), 6.9(m, 1H), 6.8-6.5 (m, 2H), 6.7-6.5 (m, 2H), 6.7 (m, 1H), 6.4 (m, 1H),5.3-5 (m, 1H), 5.1/4.7/3.6 (3*m, 1H), 5-3.6 (m, 2H), 5-3.6 (m, 2H), 4.8(m, 2H), 3.8-3.6 (m, 3H), 3.6/3.4 (m, 2H), 3.4 (m, 6H), 3.1-2.5 (m, 2H),2.9-1.9 (m, 2H), 2.5-1.7 (m, 4H), 1.8-1.4 (m, 6H), 1.6-1.3 (m, 4H)

Step G:N-(4-Hydroxyphenyl)-N-(1-methyl-2,3-dihydropyrrolo[2,3-b]pyridin-5-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxamide

0.71 g (11 mmol) of sodium cyanoborohydride is added to a solution, in20 mL of acetic acid, of the compound obtained in Step F (2.0 g, 2.2mmol). After stirring for 14 hours at ambient temperature, 0.36 g (5.5mmol) of sodium cyanoborohydride is again added, and then the reactionmixture is heated at 50° C. for 3 hours before a second addition of 0.1eq. of sodium cyanoborohydride to complete the reaction in 30 minutes at50° C. The acetic acid is evaporated off under reduced pressure, andthen the residue is taken up in dichloromethane and washed withsaturated aqueous NaHCO₃ solution, water and brine. The combined aqueousphases are extracted with dichloromethane. The resulting organic phasesare dried over sodium sulphate, filtered and concentrated under reducedpressure. The crude product obtained is purified by chromatography oversilica gel (dichloromethane/ethanol/ammonia gradient) to yield the titlecompound in the form of a meringue.

¹H NMR (500 MHz, dmso-d6, 300K) δ ppm: 9.3 (bs, 1H), 7.5/7.4/7.3 (3*m,1H), 7.2/6.7 (2*m, 1H), 7.2 (m, 1H), 7.1-6.8 (m, 4H), 6.9/6.7 (m, 1H),6.9 (m, 1H), 6.8-6.5 (m, 2H), 6.7-6.5 (m, 2H), 5.3-5.1 (2*d, 1H),5.1/4.7/3.6 (3*m, 1H), 4.9/4.2-3.5 (2*m, 1H), 4.9/4.2-3.5 (2*, 1H),4.9-4.8 (m, 2H), 3.6/3.4 (2*m, 4H), 3.4/3.3 (m, 2H), 3.4 (m, 6H),3.1-2.5 (m, 4H), 3-2.4 (m, 2H), 2.8/2.6 (m, 3H), 2.6-1.7 (m, 6H),1.9-1.3 (m, 10H)

Step H:N-(4-Hydroxyphenyl)-N-(1-methyl-2,3-dihydropyrrolo[2,3-b]pyridin-5-yl)-3-[2-[(3S)-3-(morpholinomethyl)-3,4-dihydro-1H-isoquinoline-2-carbonyl]-4-[2-oxo-2-(1-piperidyl)ethoxy]phenyl]-5,6,7,8-tetrahydroindolizine-1-carboxamidehydrochloride

The base obtained in Step G (0.60 g, 0.69 mmol) is dissolved inacetonitrile and then converted into salt form using 0.7 mL (0.7 mmol)of 1N HCl solution. The solution is filtered, frozen and thenlyophilised to provide the title compound in the form of a powder.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=68.02:68.06; % H=6.49:6.21; % N=10.89:10.87; % Cl=4.14:3.94

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₅₁H₅₇N₇O₆

[M+H]⁺ calculated: 864.4445

[M+H]⁺ measured: 864.4443

Step I:4-[(1-Methyl-2,3-dihydro-1H-pyrrolo[2,3-b]pyridin-5-yl){[3-(2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-4-[2-oxo-2-(piperidin-1-yl)ethoxy]phenyl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

IR (cm⁻¹): ν: C═O: 1625; ν: (phosphate; ether): 1229, 1138, 1115, 982;γ: >CH Ar: 880-748-745

Elemental Microanalysis:

% C % H % N Calculated 62.00 5.71 9.92 Found 61.45 5.53 9.96

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₅₁H₅₆N₇Na₂O₉P

[M−2Na+3H]⁺ calculated: 944.4106

[M−2Na+3H]⁺ measured: 944.4116

EXAMPLE 244-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate Step A:5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-N-(1-methyl-1H-pyrazol-4-yl)-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation4 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 5″. The product obtained is finally subjected toa step of conversion into salt form in the presence of HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

Elemental Microanalysis: (%, Theoretical:Measured)

% C=63.77:62.83; % H=5.63:5.83; % N=11.74:11.29; % Cl-=4.95:5.42

Step B:4-[{[5-(5-Chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

IR (cm⁻¹): ν: C═O: 1625; ν: (phosphate; ether): 1241, 1146, 1112, 983

Elemental Microanalysis:

% C % H % N Calculated 56.83 4.77 10.46 Found 56.82 4.58 10.43

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₃₈H₃₈ClN₆Na₂O₇P

[M−2Na+3H]⁺ calculated: 759.2457

[M−2Na+3H]⁺ measured: 759.2465

EXAMPLE 254-[(5-Cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}amino]phenyldisodium phosphate Step A:N-(5-Cyano-1,2-dimethyl-1H-pyrrol-3-yl)-5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-N-(4-hydroxyphenyl)-1,2-dimethyl-1H-pyrrole-3-carboxamidehydrochloride

The procedure is in accordance with a protocol analogous to thatdescribed in Steps A-D of Example 1 replacing, on the one hand, thecompound of Preparation 1 used in Step A by the compound of Preparation7 and, on the other hand, the compound of Preparation 1″ used in Step Cby that of Preparation 7″. The product obtained is finally subjected toa step of conversion into salt form in the presence of HCl in ether.After filtration and lyophilisation in a mixture of acetonitrile/water,the expected compound is obtained.

High-Resolution Mass Spectrometry (ESI/FIA/HR and MS/MS):

Empirical formula: C₄₁H₄₁FN₆O₄

[M+H]⁺ calculated: 701.3246

[M+H]⁺ measured: 701.3282

Step B:4-[(5-Cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}amino]phenyldisodium phosphate

The procedure is in accordance with a protocol analogous to thatdescribed in Steps B and C of Example 16.

³¹P NMR (400/500 MHz, CD₃OD) δ ppm: −0.5

IR (cm⁻¹): ν: —CN: 2211 cm⁻¹; ν: C═O: 1629; ν: (phosphate; ether): 1236,1114, 984

Elemental Microanalysis:

% C % H % N Calculated 59.71 4.89 10.19 Found 60.09 4.95 9.88

High-Resolution Mass Spectrometry (ESI+):

Empirical formula: C₄₁H₄₀FN₆Na₂O₇P

[M−2Na+3H]⁺ calculated: 781.2909

[M−2Na+3H]⁺ measured: 781.2898

Pharmacological and Pharmacokinetic Studies

For clarification purposes, and in anything which follows, compounds offormula (I′) will be referred to as “drug of Example x” from which theyhave been derived. As for an example,N-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide will be referred to as the “drug from Example 1”.

EXAMPLE A1 Induction of Caspase Activity in Vivo by Compounds of Formula(I′)

The ability of the compounds of formula (I′) to activate caspase 3 isevaluated in a xenograft model of RS4;11 leukaemia cells.

1×10⁷ RS4;11 cells are grafted sub-cutaneously into immunosuppressedmice (SCID strain). 25 to 30 days after the graft, the animals aretreated orally with the various compounds. Sixteen hours aftertreatment, the tumour masses are recovered and lysed, and the caspase 3activity is measured in the tumour lysates.

This enzymatic measurement is carried out by assaying the appearance ofa fluorigenic cleavage product (DEVDase activity, Promega). It isexpressed in the form of an activation factor corresponding to the ratiobetween the two caspase activities: the activity for the treated micedivided by the activity for the control mice.

N-(4-Hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide (also referred to as the drug from Example 1) was tested. Ata dose of 100 mg/kg p.o., the in vivo caspase activation factor is 29.3.

The results obtained show that the compounds of formula (I′) are capableof inducing apoptosis in RS4;11 tumour cells in vivo.

EXAMPLE A2 Quantification of the Cleaved Form of Caspase 3 in VivoBrought About by Compounds of formula (I′)

The ability of the compounds of formula (I′) to activate caspase 3 isevaluated in a xenograft model of RS4;11 leukaemia cells.

1×10⁷ RS4;11 cells are grafted sub-cutaneously into immunosuppressedmice (SCID strain). 25 to 30 days after the graft, the animals aretreated orally with the various compounds. After treatment, the tumourmasses are recovered and lysed, and the cleaved (activated) form ofcaspase 3 is quantified in the tumour lysates.

The quantification is carried out using the “Meso Scale Discovery (MSD)ELISA platform” test, which specifically assays the cleaved form ofcaspase 3. It is expressed in the form of an activation factorcorresponding to the ratio between the quantity of cleaved caspase 3 inthe treated mice divided by the quantity of cleaved caspase 3 in thecontrol mice.

The results show that the compounds of formula (I′) are capable ofinducing apoptosis in RS4;11 tumour cells in vivo.

TABLE 1 Caspase activation factors (cleaved caspase 3 MSD test in thetumours of treated mice versus control mice) in vivo, after treatment bythe oral route Dose Sampling Activation factor +/− Compound tested(mg/kg) time S.E.M. Drug from Example 13 12.5 2 hours 24.5 +/− 7.5 Drugfrom Example 19 12.5 2 hours 13.5 +/− 1.2 Drug from Example 20 12.5 2hours 52.0 +/− 8.6 Drug from Example 21 12.5 2 hours 22.6 +/− 2.4 Drugfrom Example 24 25 2 hours 45.7 +/− 2.0 Drug from Example 25 12.5 2hours  38.7 +/− 10.7 Drug from Example 15 25 2 hours 29.8 +/− 4.0

EXAMPLE A3 Quantification of the Cleaved Form of Caspase 3 in VivoBrought About by Compounds of Formula (I)

The ability of the compounds of formula (I) to activate caspase 3 isevaluated in a xenograft model of RS4;11 leukaemia cells in accordancewith the protocol given in Example A2.

TABLE 2 Caspase activation factors (cleaved caspase 3 MSD test in thetumours of treated mice versus control mice) in vivo, after treatment bythe oral route Compound Dose Sampling Activation tested (mg/kg) timefactor +/− S.E.M. Example 13 12.5 2 hours 58.6 +/− 4.6 Example 1 50 2hours 21.2 +/− 1.3 Example 19 12.5 2 hours 27.5 +/− 3.5 Example 20 12.52 hours 62.1 +/− 3.4 Example 21 25 2 hours 55.2 +/− 6.2 Example 24 25 2hours 60.5 +/− 4.5 Example 25 12.5 2 hours 61.8 +/− 8.9 Example 15 25 2hours 12.1 +/− 1.1

EXAMPLE B Solubility of Compounds of Formula (I)

The solubility of compounds of formula (I) in water was measured andcompared with that of compounds of formula (I′).

More specifically,4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate (also referred to as the compound of Example 1) wastested and compared toN-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide (also referred to as the drug from Example 1).

The solubility of the compound of Example 1 in water is greater than orequal to 10 mg/mL (12.6mM) whereas that of the associated drug is only40 μg/mL (56.2 μM). The solubilities of the compounds were also measuredin a medium buffered to physiological pH (cf. Table 3).

TABLE 3 Solubilities in aqueous medium (buffer solution: phosphate0.33M, pH = 7.4) of compounds of formula (I) and the associatedcompounds of formula (I′), measured at four concentrations: 10 μM, 20μM, 50 μM and 100 μM Solubility at Solubility Solubility SolubilityCompound tested 10 μM at 20 μM at 50 μM at 100 μM Example 19 SolubleSoluble Soluble Soluble Drug from Example 19 Soluble Soluble SolubleInsoluble Example 20 Soluble Soluble Soluble Soluble Drug from Example20 Soluble Insoluble Insoluble Insoluble Example 25 Soluble SolubleSoluble Soluble Drug from Example 25 Soluble Soluble Insoluble InsolubleExample 1 Soluble Soluble Soluble Soluble Drug from Example 1 InsolubleInsoluble Insoluble Insoluble

The results show that the compounds of formula (I) are much more solublethan the compounds of formula (I′). Only compounds of formula (I)exhibit solubilities greater than or equal to 100 μM.

EXAMPLE C In Vivo Conversion of Compounds of Formula (I)

The pharmacokinetic profile of the phosphate compounds of formula (I) isevaluated in a lipid formulation and in aqueous solution in female SCIDmice. This is compared to the pharmacokinetic profile of compounds offormula (I′) in a lipid formulation. More specifically,4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate (also referred to as the compound of Example 1) wastested and compared toN-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide (also referred to as the drug from Example 1).

Lipid Formulation of the Compound of Example 1

The compound of Example 1 is prepared in a mixture of anhydrousethanol/polyethylene glycol 300/water (10/40/50, v/v/v) intended foradministration by the p.o. route. The study is carried out in 2 groupsof SCID mice to which the compound of Example 1 is administered underthe following conditions:

-   -   Group 1: 3 mg/kg p.o. (gavage, 10 mL/kg),    -   Group 2: 25 mg/kg p.o. (gavage, 10 mL/kg).

Blood samples are taken at the following points in time (3 samples peranimal and 3 animals for each point in time): 0.25 hr, 0.5 hr, 1 hr, 2hrs, 6 hrs and 24 hrs after oral administration.

Aqueous Formulation of the Compound of Example 1

The compound of Example 1 is also administered by the oral route in anaqueous medium to SCID mice, under the following conditions:

-   -   Group 1: 30 mg/kg p.o. dissolved in 1 mM sodium carbonate        solution (gavage, 10 mL/kg),    -   Group 2: 100 mg/kg p.o. in water (gavage, 10 mL/kg).

Blood samples are taken at the following points in time (3 animals foreach point in time and 1 sample per animal): 0.25 hr, 0.5 hr, 1 hr, 2hrs, 4 hrs, 6 hrs and 24 hrs after oral administration.

For all formulations of the compound of Example 1, the blood therebycollected is centrifuged and the plasma is transferred to tubescontaining 1M hydrochloric acid. The plasma concentrations of thephosphate compound (prodrug) and its hydroxylated homologue (drug) aredetermined simultaneously using a method of liquid chromatographycoupled with mass spectrometric detection (TFC-LC-MS/MS). The limit ofdetection for both entities is 0.5 ng/mL.

Lipid Formulation of the Drug from the Compound of Example 1

The drug from Example 1 is prepared in a mixture of anhydrousethanol/polyethylene glycol 300/water (10/40/50, v/v/v) intended foradministration by the p.o. route. The study is carried out in severalgroups of SCID mice to which the drug from Example 1 is administeredunder the following conditions:

-   -   Group 1: 3 mg/kg p.o. (gavage, 10 mL/kg),    -   Group 2: 30 mg/kg p.o. (gavage, 10 mL/kg),    -   Group 3: 25 mg/kg p.o. (gavage, 10 mL/kg),    -   Group 4: 100 mg/kg p.o. (gavage, 10 mL/kg).

Blood samples are taken at the following points in time (3 animals foreach point in time and 3 samples per animal for Groups 1-2 and 1 sampleper animal for Groups 3 and 4):

-   -   p.o.: before administration and then 0.25 hr, 0.5 hr, 0.75 hr, 1        hr, 2 hrs, 4 hrs, 6 hrs, 8 hrs, 16 hrs and 24 hrs after oral        administration at 3 and 30 mg/kg,    -   p.o.: 0.5 hr, 2 hrs, 6 hrs, 16 hrs and 24 hrs after oral        administration at 25 mg/kg and 0.5 hr, 1 hr, 2 hrs, 6 hrs, 16        hrs, 30 hrs and 48 hrs after oral administration at 100 mg/kg.

The plasma from the blood samples collected after administration of thelipid formulations of the drug from the compound of Example 1 areanalysed by liquid chromatography coupled with mass spectrometricdetection. The limit of quantification of the drug from the compound ofExample 1 is less than or equal to 0.5 ng/mL.

Non-compartmental pharmacokinetic analysis is carried out on the meanvalues of the plasma concentrations of the compounds tested. The resultsare shown in Tables 4 and 5 hereinbelow.

The results show that, whatever the dose (from 3 to 100 mg/kg) and thecarrier (lipid or aqueous formulation), the major part of the prodrug offormula (I) is rapidly converted in vivo into the corresponding drug offormula (I′) (see Table 4). Plasma exposure of the prodrug (C_(max),AUC) is low in comparison to that of the corresponding drug. The resultsalso show that the plasma concentration of the drug so measured (afteradministration of the prodrug) is equivalent to or even greater thanthat measured after direct administration of the drug by the oral route(see Table 5).

TABLE 4 Compound Compound measured administered Example 1 Drug fromExample 1 Example 1, 3 C_(max) (ng/mL) = 16 C_(max) (ng/mL) = 342 mg/kgp.o. Lipid T_(max) (h) = 0.25 T_(max) (h) = 0.25 formulation AUC_(t) (ng· h/mL) = 5 AUC_(t) (ng · h/mL) = 314 Example 1, 25 C_(max) (ng/mL) =244 C_(max) (ng/mL) = 6204 mg/kg p.o. Lipid T_(max) (h) = 0.25 T_(max)(h) = 0.5 formulation AUC_(t) (ng · h/mL) = 92 AUC_(t) (ng · h/mL) =20952 Example 1, 30 C_(max) (ng/mL) = 391 C_(max) (ng/mL) = 11967 mg/kgp.o. T_(max) (h) = 1.0 T_(max) (h) = 0.5 Aqueous AUC_(t) (ng · h/mL) =879 AUC_(t) (ng · h/mL) = 49416 formulation Example 1, 100 C_(max)(ng/mL) = 359 C_(max) (ng/mL) = 28066 mg/kg p.o. T_(max) (h) = 2.0T_(max) (h) = 2.0 Aqueous AUC_(t) (ng · h/mL) = 797 AUC_(t) (ng · h/mL)= 168478 formulation

TABLE 5 Compound measured Compound administered Drug from Example 1 Drugfrom Example 1, 3 mg/kg p.o. C_(max) (ng/mL) = 295 Lipid formulationT_(max) (h) = 0.25 AUC_(t) (ng · h/mL) = 225 Drug from Example 1, 25mg/kg p.o. C_(max) (ng/mL) = 5070 Lipid formulation T_(max) (h) = 2.0AUC_(t) (ng · h/mL) = 20400 Drug from Example 1, 30 mg/kg p.o. C_(max)(ng/mL) = 8580 Lipid formulation T_(max) (h) = 1.0 AUC_(t) (ng · h/mL) =24200 Drug from Example 1, 100 mg/kg p.o. C_(max) (ng/mL) = 25878 Lipidformulation T_(max) (h) = 0.5 AUC_(t) (ng · h/mL) = 148046

More specifically, p.o. administration of the prodrug in an aqueouscarrier makes it possible to obtain plasma concentrations of the drugwhich are equivalent to or even greater than those obtained after directp.o. administration of the drug in a lipid carrier. The prodrugtherefore offers the benefit of ease of formulation compared to thecorresponding drug, especially in an aqueous medium, which is veryadvantageous with a view to clinical development. Indeed, as Example Dshows, the drug from Example 1 is difficult to formulate in an aqueousmedium.

Aqueous Formulation of the Compounds of Examples 20 and 25

The compounds of Examples 20 and 25 are administered by the oral routein an aqueous medium to SCID mice, under the following conditions:

-   -   Group 1: 3 mg/kg p.o. in solution in 1M sodium carbonate        (gavage, 10 mL/kg),    -   Group 2: 25 mg/kg p.o. in solution in 1M sodium carbonate        (gavage, 10 mL/kg).

Blood samples are taken at the following points in time (3 animals foreach point in time): 0.25 hr, 0.5 hr, 1 hr, 2 hrs, 6 hrs and 24 hrsafter oral administration.

The blood thereby collected is centrifuged and the plasma is transferredto tubes containing 1M hydrochloric acid. The plasma concentrations ofthe phosphate compound (prodrug) and its hydroxylated homologue (drug)are determined simultaneously using a method of liquid chromatographycoupled with mass spectrometric detection (TFC-LC-MS/MS). The limit ofdetection for both entities is 0.5 ng/mL.

Lipid Formulation of the Drug from the Compounds of Examples 20 and 25

The drugs from Examples 20 and 25 are prepared in a mixture ofpolyethylene glycol 300/ethanol/Phosal 50PG (30/10/60, v/v/v) intendedfor administration by the p.o. route to SCID mice, under the followingconditions:

-   -   Group 1: 3 mg/kg p.o. (gavage, 10 mL/kg),    -   Group 2: 25 mg/kg p.o. (gavage, 10 mL/kg).

Blood samples are taken at the following points in time (3 animals foreach point in time): 0.25 hr, 0.5 hr, 1 hr, 2 hrs, 6 hrs and 24 hrsafter oral administration.

The blood thereby collected is centrifuged and the plasma is transferredto tubes containing 1M hydrochloric acid. The plasma concentrations ofthe drug are determined using a method of liquid chromatography coupledwith mass spectrometric detection (TFC-LC-MS/MS). The limit ofquantification is 0.5 ng/mL.

Non-compartmental pharmacokinetic analysis is carried out. The meanresults are shown in Tables 6, 7, 8 and 9 hereinbelow.

TABLE 6 Example 20 Compound Compounds measured administered Example 20Drug from Example 20 Example 20, C_(max) (ng/mL) = BLQ C_(max) (ng/mL) =56 3 mg/kg p.o. T_(max) (h) = ND T_(max) (h) = 1.0 Aqueous AUC_(t) (ng ·h/mL) = ND AUC_(t) (ng · h/mL) = 51 formulation Example 20, C_(max)(ng/mL) = 127 C_(max) (ng/mL) = 3701 25 mg/kg p.o. T_(max) (h) = 0.25T_(max) (h) = 1.0 Aqueous AUC_(t) (ng · h/mL) = 106 AUC_(t) (ng · h/mL)= 8724 formulation ND: not determined BLQ: below the limit ofquantification

TABLE 7 Example 20 Compound measured Compound administered Drug fromExample 20 Drug from Example 20, 3 mg/kg p.o. C_(max) (ng/mL) = 39 Lipidformulation T_(max) (h) = 1.0 AUC_(t) (ng · h/mL) = 55 Drug from Example20, 25 mg/kg p.o. C_(max) (ng/mL) = 5524 Lipid formulation T_(max) (h) =2.0 AUC_(t) (ng · h/mL) = 10172

TABLE 8 Example 25 Compound Compounds measured administered Example 25Drug from Example 25 Example 25, C_(max) (ng/mL) = 17 C_(max) (ng/mL) =29 3 mg/kg p.o. T_(max) (h) = 1.0 T_(max) (h) = 1.0 Aqueous AUC_(t) (ng· h/mL) = 14 AUC_(t) (ng · h/mL) = 31 formulation Example 25, C_(max)(ng/mL) = 106 C_(max) (ng/mL) = 2232 25 mg/kg p.o.T_(max) (h) = 1.0T_(max) (h) = 1.0 Aqueous AUC_(t) (ng · h/mL) = 114 AUC_(t) (ng · h/mL)= 3965 formulation

TABLE 9 Example 25 Compound measured Compound administered Drug fromExample 25 Drug from Example 25, 3 mg/kg p.o. C_(max) (ng/mL) = 33 Lipidformulation T_(max) (h) = 1.0 AUC_(t) (ng · h/mL) = 37 Drug from Example25, 25 mg/kg p.o. C_(max) (ng/mL) = 3004 Lipid formulation T_(max) (h) =1.0 AUC_(t) (ng · h/mL) = 5704

The results show that, whatever the dose (3 or 25 mg/kg), the major partof the prodrugs of formula (I) is rapidly converted in vivo into thecorresponding drugs of formula (I′) (see Tables 6, 7, 8 and 9). Plasmaexposures of the prodrugs (C_(max), AUC) are low in comparison toexposures of the corresponding drugs. The results also show that theplasma concentrations of the drugs so measured (after administration ofthe prodrugs) are equivalent to those measured after directadministration of the drugs by the oral route (see Tables 7 and 9).

EXAMPLE D In Vivo Pharmacokinetic Profile of the Compounds of Formula(I′)

The pharmacokinetic profile ofN-(4-hydroxyphenyl)-3-{6-[((3S)-3-(4-morpholinylmethyl)-3,4-dihydro-2(1H)-isoquinolinyl)carbonyl]-1,3-benzodioxol-5-yl}-N-phenyl-5,6,7,8-tetrahydro-1-indolizinecarboxamide (also referred to as the drug from Example 1) is alsoevaluated in a lipid and aqueous formulation in the Wistar rat.

The drug from Example 1 is prepared in an aqueous suspension inhydroxyethylcellulose 1% (w/v) in water and compared to a lipidformulation composed of a mixture of anhydrous ethanol/polyethyleneglycol 400/Phosal 50PG (10/30/60, v/v/v). The two formulations areadministered by the oral route to male Wistar rats (3 rats performulation) at a dose of 100 mg/kg p.o. (gavage, 10 mL/kg).

Blood samples are taken at the following points in time from each animal(3 animals/point in time): 0.25 hr, 0.5 hr, 0.75 hr, 1 hr, 2 hrs, 4 hrs,8 hrs and 24 hrs after oral administration.

The plasma concentrations of the tested compound are determined afterextraction followed by liquid chromatography coupled with massspectrometric detection. The limit of quantification is 0.1 ng/mL. Theresults are presented in the Table hereinbelow:

TABLE 10 Compound measured Compound administered Drug from Example 1Drug from Example 1, 100 mg/kg p.o. C_(max) (ng/mL) = 816 Aqueousformulation AUC_(t) (ng · h/mL) = 3480 Drug from Example 1, 100 mg/kgp.o. C_(max) (ng/mL) = 5070 Lipid formulation AUC_(t) (ng · h/mL) =42900

The results show that the lipid formulation makes possible much betterplasma exposure of the drug from Example 1 than the aqueous formulation.

EXAMPLE E In Vitro Test on Human Caco-2 Cells

The cellular transport from A to B (Apical to Basolateral) of thephosphate compounds of formula (I) and the compounds of formula (I′)(corresponding drugs) is studied in human Caco-2 cells. Each compound isdeposited apically at 1 or 3 μM (in duplicate) and then incubated for120 minutes.

Several samples are taken during the experiment;

-   -   apically: immediately after deposition (t=0) and at 120 minutes    -   basolaterally: at the end of the experiment (120 minutes)

The concentrations of the phosphate compound (prodrug) and/or of itshydroxylated homologue (drug) are determined by liquid chromatographycoupled with mass spectrometric detection (LC-MS/MS). The limit ofquantification for both entities is 2 ng/mL.

The apparent permeability (P_(app)) and the predicted absorbed fraction(F_(abs)) in humans are calculated for the prodrug, for the drug afterincubation of the prodrug and for the drug after incubation of the drug(Hubatsch et al, Nat Protoc. 2007; 2(9), 2111-2119).

The experiment yield, which corresponds to the ratio (in percent) of thetotal amount of compound found at the end of the experiment versus thatincubated, is also calculated.

The results have been collated in Table 11. They show that the prodrugsof the compounds of formula (I) are markedly decomposed in the course ofthe experiment (experiment yields of <1.5%), thereby bringing about theformation of the associated drugs in substantial amounts.

At the end, the predicted absorbed fraction in humans for the drugsformed after incubation of the prodrugs is similar to that obtainedafter incubation of the drugs.

TABLE 11 Compound administered Compounds measured Example 1 Drug fromExample 1 Example 1 P_(app) (10⁻⁶ cm/s) = 0.01 P_(app) (10⁻⁶ cm/s) =0.83 F_(abs) (%) = ND F_(abs) (%) = 71 Yield (%) = 0 Yield (%) = 42 Drugfrom Example 1 P_(app) (10⁻⁶ cm/s) = 0.65 F_(abs) (%) = 67 Yield (%) =37 Example 4 Drug from Example 4 Example 4 P_(app) (10⁻⁶ cm/s) = 0.33P_(app) (10⁻⁶ cm/s) = 0.43 F_(abs) (%) = ND F_(abs) (%) = 69 Yield (%) =1.3 Yield (%) = 38 Drug from Example 4 P_(app) (10⁻⁶ cm/s) = 0.21F_(abs) (%) = 46 Yield (%) = 20 Example 5 Drug from Example 5 Example 5P_(app) (10⁻⁶ cm/s) = 0.26 P_(app) (10⁻⁶ cm/s) = 2.3 F_(abs) (%) = NDF_(abs) (%) = 86 Yield (%) = 1.2 Yield (%) = 78 Drug from Example 5P_(app) (10⁻⁶ cm/s) = 0.7 F_(abs) (%) = 68 Yield (%) = 34 Example 20Drug from Example 20 Example 20 P_(app) (10⁻⁶ cm/s) = 0 P_(app) (10⁻⁶cm/s) = 0.16 F_(abs) (%) = ND F_(abs) (%) = 16 Yield (%) = 0.94 Yield(%) = 100 Drug from Example 20 P_(app) (10⁻⁶ cm/s) = 0.29 F_(abs) (%) =25 Yield (%) = 91 Example 21 Drug from Example 21 Example 21 P_(app)(10⁻⁶ cm/s) = 0 P_(app) (10⁻⁶ cm/s) = 0.21 F_(abs) (%) = ND F_(abs) (%)= 19 Yield (%) = 0.83 Yield (%) = 100 Drug from Example 21 P_(app) (10⁻⁶cm/s) = 0.27 F_(abs) (%) = 24 Yield (%) = 82 Example 25 Drug fromExample 25 Example 25 P_(app) (10⁻⁶ cm/s) = 0 P_(app) (10⁻⁶ cm/s) = 0.22F_(abs) (%) = ND F_(abs) (%) = 20 Yield (%) = 33 Yield (%) = 48 Drugfrom Example 25 P_(app) (10⁻⁶ cm/s) = 0.49 F_(abs) (%) = 40 Yield (%) =100 ND: not determined

EXAMPLE F Anti-Tumour Activity in Vivo

The anti-tumour activity of the compounds of the invention is evaluatedin a xenograft model of RS4;11 leukaemia cells.

1×10⁷ RS4;11 cells are grafted sub-cutaneously into immunosuppressedmice (SCID strain). 25 to 30 days after the graft, when the tumour masshas reached about 150 mm³, the mice are treated orally with the variouscompounds in two different regimes (daily treatment for five days perweek for two weeks, or two treatments per week for two weeks). Thetumour mass is measured twice a week from the start of treatment.

The compounds of the invention have antitumour activity, via the oralroute, in the RS4;11 leukaemia model (acute lymphoblastic leukaemia).The results obtained show that the compounds of the invention arecapable of inducing significant tumour regression.

EXAMPLE G Pharmaceutical Composition: Tablets

1000 tablets containing a dose of 5 mg of a compound 5 g selected fromExamples 1 to 25 Wheat starch 20 g  Maize starch 20 g  Lactose 30 g Magnesium stearate 2 g Silica 1 g Hydroxypropylcellulose 2 g

1. A method of treating immune and auto-immune diseases in a subject inneed thereof, comprising administration of an effective amount of aphosphate compound of formula (I):

wherein: X and Y represent a carbon atom or a nitrogen atom, wherein Xand Y may not simultaneously represent two carbons atoms or two nitrogenatoms; A₁ and A₂, together with the atoms carrying them, form anoptionally substituted, aromatic or non-aromatic heterocycle Hetcomposed of 5, 6 or 7 ring members which may have, in addition to thenitrogen represented by X or by Y, from one to 3 heteroatoms selectedindependently from oxygen, sulphur and nitrogen, wherein the nitrogenatom may be substituted by a hydrogen atom, a linear or branched(C₁-C₆)alkyl group or a —C(O)—O-Alk group wherein Alk is a linear orbranched (C₁-C₆)alkyl group, or A₁ and A₂ independently of one anotherrepresent a hydrogen atom, a linear or branched (C₁-C₆)polyhaloalkyl, alinear or branched (C₁-C₆)alkyl group or a cycloalkyl; T represents ahydrogen atom, a linear or branched (C₁-C₆)alkyl group optionallysubstituted by from one to three halogen atoms, a (C₁-C₄)alkyl-NR₁R₂group, or a (C₁-C₄)alkyl-OR₆ group; R₁ and R₂ independently of oneanother represent a hydrogen atom or a linear or branched (C₁-C₆)alkylgroup, or R₁ and R₂ together with the nitrogen atom carrying them form aheterocycloalkyl; R₃ represents a linear or branched (C₁-C₆)alkyl group,a linear or branched (C₂-C₆)alkenyl group, a linear or branched(C₂-C₆)alkynyl group, a cycloalkyl group, a(C₃-C₁₀)cycloalkyl-(C₁-C₆)alkyl group wherein the alkyl moiety is linearor branched, a heterocycloalkyl group, an aryl group or a heteroarylgroup, wherein one or more of the carbon atoms of the preceding groups,or of their possible substituents, may be deuterated; R₄ represents aphenyl substituted in the para position by a group of formula—OPO(OM)(OM′), —OPO(OM)(O⁻M₁ ⁺), —OPO(O⁻M₁ ⁺)(O⁻M₂ ⁺), —OPO(O⁻)(O⁻)M₃²⁺, —OPO(OM)(O[CH₂CH₂O]_(n)CH₃), or —OPO(O⁻M₁ ⁺)(O[CH₂CH₂O]_(n)CH₃), orR₄ represents a pyrimidin-5-yl group substituted in the para position bya group of formula —OPO(O⁻M₁ ⁺)(O⁻M₂ ⁺), wherein M and M′ independentlyof one another represent a hydrogen atom, a linear or branched(C₁-C₆)alkyl group, a linear or branched (C₂-C₆)alkenyl group, a linearor branched (C₂-C₆)alkynyl group, a cycloalkyl or a heterocycloalkylboth composed of 5 or 6 ring members, while M₁ ⁺ and M₂ ⁺ independentlyof one another represent a pharmaceutically acceptable monovalentcation, and M₃ ²⁺ represents a pharmaceutically acceptable divalentcation and n is an integer from 1 to 5, wherein the phenyl group mayoptionally be substituted by one or more halogen atoms; R₅ represents ahydrogen atom, a halogen atom, a linear or branched (C₁-C₆)alkyl group,or a linear or branched (C₁-C₆)alkoxy group; R₆ represents a hydrogenatom or a linear or branched (C₁-C₆)alkyl group; R_(a), R_(b), R_(c) andR_(d), each independently of the others, represent, a hydrogen atom, alinear or branched (C₁-C₆)alkyl, a linear or branched (C₂-C₆)alkenyl, alinear or branched (C₂-C₆)alkynyl, an aryl, a heteroaryl, a halogenatom, a linear or branched (C₁-C₆)alkoxy group, a hydroxy group, alinear or branched (C₁-C₆)polyhalo alkyl group, a trifluoromethoxygroup, —NR₇R₇′, nitro, R₇—CO—(C₀-C₆)alkyl-, R₇—CO—NH—(C₀-C₆)alkyl-,NR₇R₇′—CO—(C₀-C₆)alkyl-, NR₇R₇′—CO—(C₀-C₆)alkyl-O—,R₇—SO₂—NH—(C₀-C₆)alkyl-, R₇—NH—CO—NH—(C₀-C₆)alkyl-,R₇—O—CO—NH—(C₀-C₆)alkyl-, a heterocycloalkyl group, or the substituentsof one of the pairs (R_(a),R_(b)), (R_(b),R_(c)) or (R_(c),R_(d))together with the carbon atoms carrying them form a ring composed offrom 5 to 7 ring members, which ring may have from one to 2 hetero atomsselected from oxygen and sulphur, wherein one or more carbon atoms ofthe ring defined hereinbefore may be deuterated or substituted by fromone to 3 groups selected from halogen and linear or branched(C₁-C₆)alkyl; R₇ and R₇′ independently of one another represent ahydrogen atom, a linear or branched (C₁-C₆)alkyl, a linear or branched(C₂-C₆)alkenyl, a linear or branched (C₂-C₆)alkynyl, an aryl or aheteroaryl, or R₇ and R₇′ together with nitrogen atom carrying them forma heterocycle composed of from 5 to 7 ring members, it being understoodthat: “aryl” means a phenyl, naphthyl, biphenyl or indenyl group,“heteroaryl” means any mono- or bi-cyclic group composed of from 5 to 10ring members, having at least one aromatic moiety and containing from 1to 4 hetero atoms selected from oxygen, sulphur and nitrogen (includingquaternary nitrogens), “cycloalkyl” means any mono- or bi-cyclic,non-aromatic, carbocyclic group containing from 3 to 10 ring members,“heterocycloalkyl” means any mono- or bi-cyclic, non-aromatic, condensedor spiro group composed of 3 to 10 ring members and containing from 1 to3 hetero atoms selected from oxygen, sulphur, SO, SO₂ and nitrogen,wherein the aryl, heteroaryl, cycloalkyl and heterocycloalkyl groups sodefined and the groups alkyl, alkenyl, alkynyl and alkoxy may beoptionally substituted by from 1 to 3 groups selected from optionallysubstituted, linear or branched (C₁-C₆)alkyl, (C₃-C₆)spiro, linear orbranched, optionally substituted (C₁-C₆)alkoxy, (C₁-C₆)alkyl-S—,hydroxy, oxo (or N-oxide where appropriate), nitro, cyano, —COOR′,—OCOR′, NR′R″, linear or branched (C₁-C₆)polyhaloalkyl,trifluoromethoxy, (C₁-C₆)alkylsulphonyl, halogen, optionally substitutedaryl, heteroaryl, aryloxy, arylthio, cycloalkyl, heterocycloalkyloptionally substituted by one or more halogen atoms or alkyl groups,wherein R′ and R″ independently of one another represent a hydrogen atomor an optionally substituted, linear or branched (C₁-C₆)alkyl group,wherein the Het group defined in formula (I) may be optionallysubstituted by from one to three groups selected from linear or branched(C₁-C₆)alkyl, hydroxy, linear or branched (C₁-C₆)alkoxy, NR₁′R₁″ andhalogen, wherein R₁′ and R₁″ independently of one another represent ahydrogen atom or an optionally substituted, linear or branched(C₁-C₆)alkyl group, or an enantiomer, a diastereoisomer, or an additionsalt thereof with a pharmaceutically acceptable acid or base.
 2. Themethod according to claim 1, wherein R₄ represents phenyl substituted inthe para position by a group of formula —OPO(OM)(OM′), —OPO(OM)(O⁻M₁ ⁺),—OPO(O⁻M₁ ⁺)(O⁻M₂ ⁺), —OPO(O⁻)(O⁻)M₃ ²⁺, —OPO(OM)(O[CH₂CH₂O]_(n)CH₃), or—OPO(O⁻M₁ ⁺)(O[CH₂CH₂O]_(n)CH₃), wherein M and M′ independently of oneanother represent a hydrogen atom, a linear or branched (C₁-C₆)alkylgroup, a linear or branched (C₂-C₆)alkenyl group, a linear or branched(C₂-C₆)alkynyl group, a cycloalkyl or a heterocycloalkyl both composedof 5 or 6 ring members, and wherein M₁ ⁺ and M₂ ⁺ independently of oneanother represent a pharmaceutically acceptable monovalent cation, andM₃ ²⁺ represents a pharmaceutically acceptable divalent cation and n isan integer from 1 to 5, wherein the phenyl group may optionally besubstituted by one or more halogen atoms.
 3. The method according toclaim 1, wherein R₄ represents phenyl substituted in the para positionby a group of formula —OPO(O⁻Na⁺)(O⁻Na⁺).
 4. The method according toclaim 1, wherein X represents a carbon atom and Y represents a nitrogenatom.
 5. The method according to claim 1, wherein the group:

represents a 5,6,7,8-tetrahydroindolizine, an indolizine or adimethylated pyrrole.
 6. The method according to claim 1, wherein Trepresents a methyl, (morpholin-4-yl)methyl or 3-(morpholin-4-yl)propylgroup.
 7. The method according to claim 1, wherein R_(a) and R_(d) eachrepresent a hydrogen atom and (R_(b),R_(c)), together with the carbonatoms carrying them, form a 1,3-dioxolane group or a 1,4-dioxane group;or R_(a), R_(c) and R_(d) each represent a hydrogen atom and R_(b)represents a hydrogen atom or a halogen atom.
 8. The method according toclaim 1, wherein R_(a) and R_(d) each represent a hydrogen atom, R_(b)represents a halogen atom and R_(c) represents a methoxy group.
 9. Themethod according to claim 1, wherein R_(a), R_(b) and R_(d) eachrepresent a hydrogen atom and R_(c) represents aNR₇R₇′—CO—(C₀-C₆)alkyl-O— group.
 10. The method according to claim 1,wherein R₃ represents a group selected from phenyl, 1H-indole,1H-pyrrolo[2,3-b]pyridine, pyridine, 1H-pyrazole, 1H-pyrrole and2,3-dihydro-1H-pyrrolo[2,3-b]pyridine, those groups optionally havingone or more substituents selected from linear or branched (C₁-C₆)alkyl,cyano and trideuteriomethyl.
 11. The method according to claim 1,wherein the compound of formula (I) is selected from:4-[{[3-(6-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}-1,3-benzodioxol-5-yl)-5,6,7,8-tetrahydroindolizin-1-yl]carbonyl}(phenyl)amino]phenyldisodium phosphate,4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(pyridin-4-yl)amino]phenyldisodium phosphate,4-({[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}[1-(trideuteriomethyl)-1H-pyrazol-4-yl]amino)phenyldisodium phosphate,4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl)amino]phenyldisodium phosphate,4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(5-cyano-1-methyl-1H-pyrrol-3-yl)amino]phenyldisodium phosphate,4-[{[5-(5-chloro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate,4-[(5-cyano-1,2-dimethyl-1H-pyrrol-3-yl){[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}amino]phenyldisodium phosphate, and4-[{[5-(5-fluoro-2-{[(3S)-3-(morpholin-4-ylmethyl)-3,4-dihydroisoquinolin-2(1H)-yl]carbonyl}phenyl)-1,2-dimethyl-1H-pyrrol-3-yl]carbonyl}(1-methyl-1H-pyrazol-4-yl)amino]phenyldisodium phosphate, or an enantiomer, a diastereoisomer, or an additionsalt thereof with a pharmaceutically acceptable acid or base.
 12. Themethod according to claim 1, wherein the compound of formula (I) isadministered as a pharmaceutical composition comprising the compound offormula (I) in combination with one or more pharmaceutically acceptableexcipients.